ToRCH & Childhood Diseases

ToRCH & Childhood Diseases Reagents for Assay Development ISO Certified

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Expertise in Infectious Disease Meridian Life Science

KEY PRODUCTS FOR INFECTIOUS DISEASE

ToRCH • Toxo • Rubella • CMV • HSV-1, 2

TROPICAL • Nipah • Dengue 1, 2, 3, 4 • Chikungunya

Infectious diseases are a leading cause of death, accounting for 30% of the estimated 68 million deaths per year worldwide. They are caused by pathologic agents including bacteria, viruses, fungi and parasites. Many infectious diseases are preventable and controllable if they are accurately diagnosed and treated in a timely manner. Pregnant woman and children are at a greater risk of acquiring infectious diseases because their immune systems are not fully functioning. Pregnancy weakens the immune system, leaving expectant mothers and their fetuses more vulnerable to contracting a disease. Furthermore, babies do not start producing antibodies until they are 6 months old and a child’s immune system is not fully mature until the age of 14. In addition, children, especially under the age of 5, tend to have poor personal hygiene, increasing the spread of infectious agents. Vaccinations have helped prevent and eradicate some of the most contagious diseases, however there are a number of diseases for which researchers are still developing vaccines and many people choose to remain unvaccinated. Preventable infectious diseases still account for two thirds of child deaths worldwide. The ability to quickly diagnose the cause of an infectious disease has had a large and favorable impact on the care of pregnant women and children. Diagnostic assays that directly identify an infectious agent have become increasingly essential and new diagnostic platforms have increased the potential to detect a wider range of established and newly discovered viruses with greater sensitivity. For over 40 years, Meridian Life Science, Inc. has provided antibodies and antigens for research and commercial assay development. With a specialty in infectious disease, the company offers over 1,250 different antibodies and antigens to infectious disease markers and toxins.

• Malaria • Chagas

RESPIRATORY • RSV • Influenza A,B • Parainfluenza •  Mycoplasma pneumoniae •  Chlamydia pneumoniae •  Legionella pneumophila •  Mycobacterium tuberculosis •  Streptococcus •  Staphylococcus • SARS Coronavirus • Adenovirus

• Leishmaniasis • Leptospirosis • Japanese

GASTROINTESTINAL • Norovirus • Astrovirus • Adenovirus • Rotavirus •  Clostridium difficile Encephalitis Virus • Newcastle disease • Yellow Fever • Zika • Lyme disease • Ebola •  Cryptosporidium •  Campylobacter •  E. coli •  Salmonella •  Giardia Iamblia •  H. pylori FOOD & WATER • Hepatitis A •  Campylobacter jejuni •  E. coli •  Legionella •  Salmonella •  Shigella •  Bacillus anthracis •  Clostridium •  Listeria

CHILDHOOD • Mumps • Rubeola • EBV • Coxsackie • Rotavirus • RSV • Parvo B19 • VZV

STDs • HAV • HBV • HCV

• HSV-1, 2 • HIV-1, 2 • HPV • Chlamydia • Neisseria • Syphilis

•  Streptococcus •  Staphylococcus •  Giardia •  Cryptosporidium

ToRCH & Childhood Diseases - Reagents for Assay Development 2

Catalog Guide

Introduction...........................................................................................2 ToRCH.....................................................................................................6 Toxoplasma...........................................................................................8 Rubella................................................................................................. 10 CMV.......................................................................................................12 HSV-1 & HSV-2............................................................................... 16 Varicella zoster virus (VZV).......................................................... 18 Epstein-Barr Virus (EBV)................................................................ 20 Mumps.................................................................................................22 Rubeola (Measles)........................................................................... 23

Parvovirus B19.................................................................................. 24 Enterovirus.......................................................................................... 26 Coxsackie............................................................................................ 28 Rotavirus..............................................................................................30 Respiratory Syncytial Virus (RSV)............................................. 32 Increasing Assay Sensitivity........................................................34 Complimentary Assay Reagents...............................................35 Abbreviations....................................................................................36 Product list.........................................................................................36

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Company Overview

Meridian Life Science, Inc. is a leading large scale manufacturer of: • Antibodies • Viral antigens • Recombinant proteins • PCR enzymes • Nucleotides • Critical assay reagents Meridian has been providing innovative life science solutions and building trusted partnerships for over 40 years. Meridian’s focus is to offer products and services that help to advance the development of diagnostic assays and vaccine development. • C ommercial scale manufacturing of antigens and antibodies with protein purification expertise • F ull line of immunoassay reagents, including antigens, antibodies and blockers

• L arge scale production of reagents for molecular assays • Technical support with assay development experience • D edicated R&D and manufacturing teams • R obust and mature Quality System

ISO certified

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Extensive Capabilities and Services Immunodiagnostics • Antigens & Antibodies • Recombinant Proteins • Blocking reagents Molecular Diagnostics • Nucleotides • enzymes • qPCR/PCR reagents • NGS reagents Contract Services • Antigens & Antibodies • Cell & Viral Banking • PCR/qPCR Assay Development

Global Presence

MERIDIAN BIOSCIENCE, INC. Parent Company | Founded in 1977 | Nasdaq: VIVO Headquartered in Cincinnati, OH | 750+ Employees | Presence in 70+ Countries.

North America

EMEA

Asia Pacific

SYDNEY, AUSTRALIA Warehouse & Sales BEIJING, CHINA Wholly Owned Subsidiary

MEMPHIS, TN Viral Antigens Recombinant Proteins In Vitro Antibodies PCR Reagents BILLERICA, MA

LONDON, UK PCR Manufacturing & Sales PCR /qPCR Molecular Reagents LUCKENWALDE, GERMANY Large Scale Nucleotides PCR Enzymes Manufacturing PARIS, FRANCE EU Diagnostics Sales & Admin WATERLOO, BELGIUM EU Diagnostics Sales & Admin MILAN, ITALY EU Diagnostics Sales & Admin MODI’IN, ISRAEL BreathID ® Breath Test Systems

Magellan, Leadcare BOCA RATON, FL In Vivo Antibodies QUEBEC, CANADA GenePOC, Molecular Diagnostics

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ToRCH

ToRCH is an acronym for a group of infections that can cause significant birth defects and fetal death. Meridian Life Science offers a range of reagents which are suitable for multiple assay formats, including IgM and IgG antibody detection, rapid anti-IgM assays, and Immunofluorescence assays (IFA).

A ToRCH test measures antibodies against five groups of chronic infections: • Toxoplasmosis • Rubella • Cytomegalovirus (CMV) • Herpes simplex virus (HSV-1/2) •  Other infections (usually syphilis, hepatitis B, coxsackie virus, Epstein-Barr virus, varicella-zoster virus, and human parvovirus) These infectious diseases are all associated with congenital abnormalities resulting from maternal infection. Although the organisms typically cause only asymptomatic or mild infection to the mother, they can have much more serious consequences to the fetus. If the infection occurs during the first three months of pregnancy and if it is a primary infection (newly acquired during pregnancy), the risk of congenital abnormalities is much higher compared to a secondary or reactivated infection. CMV, which is the most common cause of congenital infectious disease, also has a much higher rate of transmission from mothers with a primary infection (10%) compared to a reactivation (1%). An important part of prenatal care is to recognize these infections in the mother and the fetus and provide suitable care.

Although these organisms typically cause only asymptomatic or mild infection in the mother, they can have serious consequences to the fetus.

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For most ToRCH organisms, the initial screening test is based on the detection of antibodies to the organism. Subsequent screening, if required, is carried out using a monoclonal antibody-based immunofluorescent assay (IFA). Assays are commercially available for the detection of IgG, IgM, or both IgG and IgM antibodies. In most cases, IgG reactivity in the absence of IgM reactivity is indicative of a past infection, while IgM reactivity in the absence of IgG reactivity indicates a current infection. However, for some ToRCH diseases such as toxoplasmosis and CMV, IgG avidity has recently been found to be useful for identifying primary infections. An IgG antibody produced in the first few months following an initial infection has a lower avidity than an IgG antibody produced several months or years later. Consequently, low-avidity antibody can be used to specifically identify high-risk mothers with a primary infection. To protect a fetus from ToRCH infection, early diagnosis through first trimester screening is critical.

TORCH IGG & IGM ANTIBODY SCREENING ASSAYS

A ToRCH serologic test must detect both IgM and IgG antibodies to the ToRCH panel of infectious diseases (Toxo, Rubella, CMV and HSV). IgM is the immediate antibody that is produced once a human is exposed to a bacteria, virus or a toxin and disappears within 2 to 3 weeks. It is then replaced by IgG which lasts for life and provides lasting immunity. All of Meridian’s ToRCH antigens are suitable for commercial IgG and/or IgM detection. They can be used in a range of immunoassay formats including, but not limited to, ELISA, LF, CLIA, rapid assays, and bead-based assays.

TORCH RAPID IGM CAPTURE ASSAYS

Rapid IgM capture assays are particularly sensitive in detecting IgM responses early in illness. The assay works by binding IgM antibodies in the patient’s specimen to a solid phase coated with an anti-IgM capture antibody. Soluble antigen is added in excess allowing the IgM antibody-antigen reaction to occur in the absence of competing IgG. Finally, a labelled detection antibody is added that has specific reactivity against the antigen. Assay sensitivity can be highly dependent on the purity of the antigen used.

TORCH IMMUNOFLUORESCENT ANTIBODY ASSAYS (IFA)

IFA is a traditional laboratory technique that utilizes fluorescent dyes to identify the presence of antibodies bound to specific antigens. Definitive diagnosis of a ToRCH infection, in particular HSV-1 or HSV-2, sometimes requires confirmation by IFA after a positive EIA result.

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Toxoplasma

Toxoplasmosis is caused by the protozoan parasite Toxoplasma gondii. It can infect most species of warm blooded animals, including humans, and can cause toxoplasmosis disease.

LIFE CYCLE OF T.GONDII

The only known definitive hosts for Toxoplasma gondii are members of family Felidae (domestic cats and their relatives). Unsporulated oocysts are shed in the cat’s feces and these take 1-5 days to sporulate in the environment and become infectious. Intermediate hosts in nature (including birds and rodents) become infected after ingesting soil, water or plant material contaminated with oocysts. Cats become infected after consuming intermediate hosts harboring tissue cysts. Humans can become infected by any of the following routes: •  Eating undercooked meat of animals harboring tissue cysts •  Consuming food or water contaminated with cat feces or by contaminated environmental samples (such as fecal- contaminated soil or changing the litter box of a pet cat) •  Blood transfusion or organ transplantation •  Transplacentally from mother to fetus

Up to a third of the world’s human population is estimated to have been infected with toxoplasma gondii . Infection is usually asymptomatic, but during the first few weeks after exposure the infection may cause a mild, flu-like illness. However, in those with weakened immune systems, such as those with AIDS and pregnant women, it can cause a serious and sometimes fatal illness.

CONGENITAL TOXOPLASMOSIS

DIAGNOSIS Diagnosis is usually made by detection of Toxo-specific IgG and IgM antibodies. A test that only measures IgG is used to determine if a person has previously been infected. When it is necessary to try to estimate the time of infection, such as in pregnancy, an IgM test is also used along with other tests such as an IgG avidity test. Diagnosis can also be made by direct observation of the parasite in stained tissue sections, cerebrospinal fluid (CSF), or other biopsy material. Congenital toxoplasmosis is a group of symptoms that occur when a fetus is infected with T. gondii. If a mother becomes infected while pregnant, the parasite can spread to a developing fetus across the placenta. The risk of congenital disease is lowest (10 – 25%) when maternal infection occurs during the first trimester and highest (60 – 90%) when maternal infection occurs during the third trimester. Congenital disease is most severe when infection is acquired in the first trimester. The overall risk of congenital infection from acute T. gondii infection during pregnancy ranges from approximately 20 – 50%.

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REAGENTS FOR SEROLOGY TESTING

8158

T. gondii Native Antigen •  Purified whole tachyzoite preparation, RH strain •  Propagated in Vero cells •  Protein concentration: ~0.5 mg/mL by BCA Assay •  Buffer: PBS, pH 7.4 containing 1% Triton x-100 T. gondii Native Antigen, Concentrated •  Purified whole tachyzoite preparation, RH strain •  Propagated in Vero cells •  High protein concentration (~1mg/ml) •  Buffer: PBS, pH 7.4 containing 1% Triton x-100

IgG & IgM Detection for ELISA and WB Assays

8200

8159

T. gondii Native Antigen, IgM •  Purified membrane fraction (highly specific for Toxoplasma p30), RH strain •  Protein concentration ~ 0.5 mg/mL (BCA Assay) •  Buffer: Phosphate Buffered Saline, pH 7.4 containing 2% N-octyl- -D-Glucopyranoside (OGP). T. gondii Chimeric Recombinant Antigen •  Contains immunogenic regions of the Toxoplamsa protein •  Produced in P. pastoris and contains a His tag at the N-terminus •  Buffer: 20 mM phosphate pH 7, 0.1 M KCL

IgM Detection for ELISA and WB Assays

R01794

IgG & IgM Detection for ELISA, CLIA and Lateral Flow

R01573

T. gondii p30 (SAG1) Recombinant Antigen •  Recombinant protein (E. coli) , ≥ 95% pure (SDS-PAGE)

•  Contains a His-tag at the N-terminus •  Calculated molecular weight 36.6kDa •  Protein concentration: ~0.8mg/ml •  Buffer: 20 mM Phosphate Buffer, pH 8.0, 1 M Sodium Chloride, 0.1% Polyoxyethylene (10) Tridecyl Ether

IgG Detection for EIA Assays

T. gondii p35 (GRA8) Recombinant Antigen •  Recombinant protein (E. coli) , ≥ 95% pure (SDS-PAGE)

R01581

IgG & IgM Detection for EIA Assays

•  Contains a His-tag at the N-terminus •  Calculated molecular weight 21.7kDa •  Protein concentration: ~1.0mg/ml •  Buffer: 20 mM Phosphate Buffer, pH 8.5, 150 mM Sodium Chloride

MAb to T. gondii •  Reacts with p30 membrane protein MAb to T. gondii , 38kDa protein •  Recognizes a 38 kDa protein MAb to T. gondii SAG1 (p30) protein •  Specific for the SAG1 protein

C86319M

C01589M

IFA Detection

C01523M

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Rubella

Rubella, also known as German Measles, is a viral illness caused by a togavirus of the genus Rubivirus and is an infection that primarily affects the skin and lymph nodes. It is generally a mild disease in adults and children, but it can have devastating effects on infants.

The primary medical danger of rubella is the infection of pregnant women, because it can cause congenital rubella syndrome (CRS) in developing fetus. Since the introduction of the measles-mumps-rubella (MMR) vaccine, rubella is much less common, however, several countries still do not include this vaccine in their immunization schedule. In the absence of vaccination, rubella is an endemic disease with epidemics every 6 to 9 years. Rubella is spread by coughing and sneezing. The virus resides in the nose and throat of an infected person with an average incubation period of 14-21 days. A person infected with rubella may spread the disease to others beginning one week before the rash occurs. Symptoms generally occur 2-3 weeks after exposure to the virus and includes a mild fever, headache and runny nose.

COUNTRIES USING RUBELLA VACCINE IN NATIONAL IMMUNIZATION SCHEDULE, 2012

Source: WHO/UNICEF coverage estimates 2012 revision, July 2013. 194 WHO Member States. Map production: Immunization Vaccines and Biologicals, (IVB). World Health Organization Date of slide: 24 July 2013.

CONGENITAL RUBELLA SYNDROME (CRS)

DIAGNOSIS Rubella virus can be detected from nasal, throat, urine, and blood specimens from infected individuals. Diagnosis is usually made by the detection of Rubella-specific IgM antibodies which are usually present 4–30 days after the onset of illness. However, reliable serodiagnosis requires the discrimination of specific IgM primary rubella from persistent, reactivated or non-specific IgM reactivity. Recent infection can be confirmed or excluded by additional assays such as Rubella IgG avidity and immunoblot analysis. A rubella infection just before conception (0-28 days) or during the first trimester in pregnancy has the highest rate of transmission to the fetus (90%) resulting in congenital rubella syndrome (CRS). At 14 weeks, this incidence is reduced to 52%, and by the end of the second trimester, the incidence drops to 25%. Fetal rubella infection often results in spontaneous abortion or severe fetal defects, including heart, brain, ear or eye malformations, deafness, microcephaly, and mental retardation. Before the introduction of the vaccine, up to 4 babies in every 1000 live births were born with CRS.

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ToRCH & Childhood Diseases - Reagents for Assay Development

REAGENTS FOR SEROLOGY TESTING

6075

Rubella Native Antigen, Grade III • Strain HPV77, propagated in Vero cells • Purified antigen

IgG Detection for ELISA & CLIA Assays

• Protein concentration: 0.1-0.2mg/mL by OD 260/280nm • Buffer: 10mM Tris, 150mM NaCl, 1mM EDTA, pH 8.0-8.4

Rubella Native Antigen, Grade IV • Strain HPV77, propagated in Vero cells • Highly purified antigen

6076

IgG & IgM Detection for ELISA, CLIA and Bead-based Assays

• Protein concentration: 0.5mg/mL by OD 260/280nm • Buffer: 10mM Tris, 150mM NaCl, 1nM EDTA, pH 8.0-8.4

6123

Rubella Native Antigen, Grade IV (PBS) • Strain HPV77, propagated in Vero cells • Highly purified antigen • Buffer: Sucrose/PBS, pH 7.4-7.7

6200

Rubella Native Antigen, RSVP TM (Capsid Free) • Strain HPV77, propagated in Vero cells • Purified using a proprietary process • Contains the envelope spike protein comprised of E1 and E2 in their native configuration • Buffer: 0.1M Na 2 CO 3 , 0.1M NaCl, pH 7.9-8.3

Very sensitive IgM Detection for ELISA and Bead- based Assays

EV9525

MAb to Rubella E1 Protein • Specific for the E1 protein

MAb to Rubella E1 Protein • Specific for E1 protein and reacts with non-reduced antigens on Western Blot • Exhibits low level of HI activity • Exhibits neutralizing activity

EV9526

Paired Ab/Ag for IgM Capture Assays

Rubella Native Antigen, Grade IV 6076

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CMV

The CMV diagnostics market is growing due to an increased prevalence of CMV infections worldwide and better disease awareness. In the U.S., 60% of the population is carrying CMV and more than 90% of those are in a high risk category (AIDS patients and prenatal babies of CMV infected mothers).

Human cytomegalovirus (CMV, also called Human herpesvirus 5) is a member of the herpes virus family and shares the characteristic ability to remain latent within the body for life within an infected individual. Although a CMV infection is typically asymptomatic in healthy persons, immunocompromised individuals such as AIDS patients, organ transplant recipients and newborn infants, are at high-risk of developing life-threatening complications from primary infections and reactivations. CMV is not considered highly contagious and the virus is generally passed through direct contact with body fluids, such as urine, saliva, breast milk, transplanted organs and blood transfusions. Healthy pregnant women are not at special risk for disease from CMV infection but between 5-8% are infected for the first time during their pregnancy, and this can lead to serious complications. Among infants born with CMV infection (congenital CMV), about 20%

Source: cdc.gov

will have permanent disabilities. There is no vaccine available to protect against CMV and public health measures focus on reducing the risk of CMV transmission to pregnant women, women of childbearing age and other people at risk of more serious infections.

CONGENITAL CMV

Congenital Cytomegalovirus (CMV) refers to a group of symptoms that occur when an infant is infected before birth and it is the most common cause of congenital viral infections worldwide. Only 10% of congenitally infected newborns display abnormalities at birth, however 80% - 90% will develop complications within the first few years of life. Symptoms of congenital CMV include hearing loss, vision impairment, and varying degrees of mental retardation. The risks for a fetus becoming infected by CMV appear to be almost exclusively associated with women who are having a primary infection during pregnancy. There appears to be little risk of CMV related complications for women who have been infected at least 6 months prior to conception. DIAGNOSIS Various diagnostic tests have been developed to detect a CMV infection including viral culture, serological assays, PCR analysis and cytopathology. The pp65 antigenemia test, in which a monoclonal antibody against CMV pp65 is used to detect a major CMV matrix protein (pp65) in leukoctyes, has the longest history in clinical use. However, it has been criticized for its subjectivity in reading positive results, time consuming and intricate procedures, difficulty in standardization, and a need for sufficient leukocytes. The ELISA IgG/IgM assay which measures antibodies to CMV, specifically CMV IgM, IgG and IgG avidity, has become the most commonly available serologic test. The detection of IgM is indicative of an acute or primary infection whereas the detection of IgG is indicative of a past infection. In the case where both IgM and IgG can be detected, the level of IgG avidity can help distinguish between an acute infection and a past infection. For this reason, newer assays have begun to incorporate the detection of anti-CMV IgM together with determination of the avidity index of anti-CMV IgG. To improve the sensitivity and specificity of CMV antibody detection, immunogenic CMV proteins have been studied and characterized during the past two decades and over 15 structural polypeptides have been identified in a natural infection. The combination of antigens selected is the most critical element affecting assay sensitivity and specificity.

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ToRCH & Childhood Diseases - Reagents for Assay Development

The most suitable proteins are reportedly: • CMV pp150: a tegument phosphoprotein detectable during both latent and re-activated infections. During primary infection the antibody response to pp150 may be delayed. • CMV pp52: the major DNA binding protein, nonstructural nuclear phosphoprotein which is regarded as an early marker of seroconversion • CMV pp65: major structural phosphoprotein (lower matrix) and main component of extracellular virus particles. The antibody response is detectable during early infection only • CMV gB Antigen: a membrane glycoprotein which is the most abundant component of the viral envelope and a target of neutralizing antibodies • CMV pp38: structural protein suggested to be an important immunodominant protein in early infection Evidence also suggests that CMV-IgM detection against viral structural proteins (pp150 and pp38) are a valuable parameter for the early diagnosis of a recurrent CMV infection. Several diagnostic manufacturers have incorporated a combination of CMV lysate and CMV recombinant proteins to improve assay performance. REAGENTS FOR SEROLOGY TESTING

7504

CMV-G Native Antigen • Whole cell extract, >10% viral protein • Strain AD169 propagated in human fibroblast cells

• 0.25 mg/mL by OD 260/280nm • Buffer: 0.1M Glycine, pH 9.3-9.7

7517

CMV Ext-2 Native Antigen (Concentrate) • Enriched for cell surface glycoprotein antigens, >10% viral protein • Strain AD169 propagated in human fibroblast cells • Protein concentration: 0.2-1mg mg/mL by OD 260/280nm • Buffer: 0.1M Glycine, pH 9.5±0.2 CMV Native Antigen • Viral lysate prepared by centrifugation to remove cell debris • Strain AD169 propagated in MRC-5 cells • Buffer: Glycine buffered saline, pH 9.5 CMV gB Native Antigen • Strain AD169 propagated in human fibroblast cells • Buffer: 0.1M Glycine, pH 9.5 ± 0.2 5 CMV gB Recombinant Antigen • Contains the CMV gB immunodominant region (Strain C194) and a GST fusion partner • Immunoreactive with CMV positive sera • Produced in E. coli , >95% pure (SDS-PAGE) • Buffer: 25mM Tris-HCl, 1mM EDTA, pH 7.2 and 50% glycerol

7600

IgG Detection for EIA Assays

EV7509

R18102

R01686

CMV IgM Native Antigen (Concentrate) • Preparation of nuclear extract and ER antigens • Strain AD169 propagated in human fibroblast cells • Buffer: 0.1M Glycine, pH 9.3-9.7 CMV Chimeric Recombinant Antigen • Produced in E. coli , >95% pure (SDS-PAGE) • 31.6kDa calculated MW • Buffer: 20 mM NaP @ pH8, 1 M NaCl

7511

IgM Detection for EIA Assays

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CMV continued

REAGENTS FOR SEROLOGY TESTING (cont)

EV9268

CMV II Native Antigen • Purified surface extract containing pp65, pp52, pp150 and other key proteins • Strain AD169 propagated in human fibroblast cells • Buffer: 0.1M Glycine, pH 9.5±0.2

7507

CMV III Native Antigen • Purified surface extract enriched for pp65, >60% viral protein • Strain AD169 propagated in human fibroblast cells • Protein concentration: 0.8-1.2mg/mL by BCA • Buffer: 0.1M Glycine, pH 9.3-9.7 CMV Native Antigen • Viral lysate prepared by centrifugation to remove cell debris

IgG & IgM Detection for EIA Assays

7600

• Strain AD169 propagated in MRC-5 cells • Buffer: Glycine buffered saline, pH 9.5

R01763

Coxsackievirus A16 Recombinant Antigen • Enterovirus A • Produced in E. coli, contains a GST fusion partner • Buffer: Tris buffer with salts, pH 8.0, 0.09% Sodium Azide

Suitable for use as a Control or for Antibody Detection in Lateral FlowAssays

CMV pp52 (UL44) Recombinant Antigens

R01565

• Represents a truncated portion of CMV pp52 gene fused to a GST-tag • Molecular weight of 44kDa • Produced in E. coli , ≥ 95% pure (PAGE) • Buffer: 50mM Tris-HCl, pH 7.2 containing 60mM NaCl, 10mM GSH, 50% Glycerol • Represents a truncated portion of CMV pp52 gene fused to a His-tag at the N-terminus • Produced in E. coli , ≥ 95% pure (SDS-PAGE) • Buffer: 20mM Sodium Phosphate, 1M NaCl, pH 8.0 • Represents residues 202-434 of CMV pp52, fused with a GST tag at the N-terminus • Molecular weight of 51kDa • Produced in E. coli , ≥ 90% pure (SDS-PAGE) • Buffer: 50mM Tris-HCl, 60mM NaCl, 10mM GSH, 0.25% Sarcosyl, 50% Glycerol, pH 8.0 • Represents CMV pp65 (strain AD169) gene fused to a GST-tag • Molecular weight of 50kDa • Produced in E. coli , ≥ 95% pure (SDS-PAGE) • Buffer: 25mM Tris-HCl, 1mM EDTA, pH 7.2 containing 50% Glycerol • Represents CMV pp65 gene fused to a His-tag at the N-terminus • Molecular weight of 35kDa • Produced in E. coli , ≥ 95% pure (SDS-PAGE) • Buffer: 20mM Sodium Phosphate, 1M NaCl, 0.1% Polyoxyethylene (10) Tridecyl Ether and 8M Urea, pH 8.0

R01561

R18062

IgM Detection for EIA Assays

R01562

R18412

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CMV pp38 (UL80a) Recombinant Antigen

R18512

• Represents the immunodominant regions of the CMV pp38 gene • Molecular weight of 55kDa • Produced in E. coli , ≥ 95% pure (PAGE) • Buffer: 25mM Tris-HCl, 1mM EDTA, pH 7.2 containing 50% Glycerol

IgM Detection for EIA Assays

CMV pp150 (UL32) Recombinant Antigen

R01564

• Represents residues 1011-1048 of CMV pp150, fused with a GST tag • Molecular weight of 35kDa • Produced in E. coli , ≥ 90% pure (PAGE) • Buffer: 50mM Tris-HCl, 60mM NaCl, 10mM GSH, 0.25% Sarcosyl, 50% Glycerol, pH 8.0 • Represents a truncated portion of CMV pp150 gene fused to a His-tag at the N-terminus • Molecular weight of 16kDa • Produced in E. coli , ≥ 95% pure (PAGE) • Buffer: 20mM sodum phospaste, 1M NaCl, 0.1% Polyoxyethylene (10) Tridecyl Ether, pH 7.5. • Contains the CMV pp150 immunodominant region and a GST tag • Molecular weight of 32kDa • Produced in E. coli , ≥ 90% pure (PAGE) • Buffer: 25 mM Tris-HCl, pH 8.0, 1 mM EDTA containing 50% Glycerol

R01563

IgG & IgM Detection for EIA Assays

R18113

C86314M

MAb to CMV Early Antigen • Reacts with 65kDa protein of CMV

Paired Ab/Ag for IgM Capture Assays

CMV II Native Antigen EV9268

C8A022M

MAb to CMV Immediate Early Antigen (IEA) • Specific for CMV IEA pp72

IFA Detection

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HSV-1 & HSV-2

Herpes simplex virus (HSV) types 1 and 2 are common infections worldwide. However, the majority of infected individuals remain undiagnosed because they are asymptomatic.

HSV-1 is usually transmitted during childhood through contact with oral secretions (cold sores). Seroprevalence studies indicate about 60% of adults in the United States are infected with this virus. HSV-2 is usually spread by sexual contact (genital herpes). Consequently this infection usually occurs later in life and the seroprevalence rates vary dramatically by geographic region. Both HSV-1 and HSV-2 establish a lifelong, latent infection in the nervous system and there is no cure. Antiviral medications can reduce the frequency, duration and severity of outbreaks and over a period of several years, many infected individuals experience less severe symptoms and fewer outbreaks, although they are still contagious to others. The greatest risk of an HSV infection is in neonates and infants, when an infected mother passes it to her fetus in utero or during delivery. A neonatal HSV infection can be devastating to an infant and 70 - 85% of these infections are caused by HSV-2. Many infants infected with HSV are born prematurely and approximately 4% can develop congenital HSV which has serious consequences including death.

HERPES SIMPLEX VIRUS

CONGENITAL HERPES SIMPLEX

DIAGNOSIS Mother and baby are usually tested simultaneously if herpes is suspected. Diagnostic methods include serological tests such as ELISA and IFA, as well as PCR blood tests and cell culture. Due to a high degree of genetic similarity between HSV-1 and HSV-2, most viral proteins induce a cross-reactive antibody response that hampers the discrimination between HSV-1 and HSV-2 infections using serological approaches. Since the discovery of the serologically distinct HSV viral envelope glycoproteins gG-1 (HSV-1) and gG-2 (HSV-2), new type-specific immunoassays have been developed that are capable of discriminating between HSV-1 and HSV-2 infections. Since antibodies may take several weeks to reach detectable levels after primary infection, negative results should be confirmed by repeat testing 4 to 6 weeks after a suspected early infection. In most cases, babies contract congenital herpes in the birth canal during delivery (especially if the mother has an active outbreak of genital herpes at the time of delivery). In rare circumstances, it is possible to be infected in the uterus or immediately after birth (from being kissed or having other contact with someone who has herpes mouth sores). Congenital HSV is a serious condition and affects about 1 out of every 3,000-20,000 live births. Detection and prevention is difficult because the infected mother is typically asymptomatic. Congenital herpes symptoms usually appear within the first month of the infant’s life and antiviral treatments such as vidarabine and acyclovir have proven helpful to reduce the severity of the disease. However, infants with systemic herpes or encephalitis often do poorly, despite antiviral medications and early treatment.

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REAGENTS FOR SEROLOGY TESTING

HSV-1 Native Antigen 7305

HSV-1 Native Antigen (Concentrate) • Strain F produced in Vero cells • >10% viral protein • Buffer: 0.1M Glycine, pH 9.5 ± 0.2

7309

IgG Detection for ELISA and CLIA Assays

HSV-2 Native Antigen 7705

7749

HSV-2 Native Antigen (Concentrate) • Strain G produced in Vero cells • >10% viral protein • Buffer: 0.1M Glycine, pH 9.5 ± 0.2

VTI520

HSV-1 Recombinant Antigen, Glycoprotein G 1 • Represents amino terminal Met1-Asp190 and fused with superoxide dismutase 1 (SOD) • Produced in Saccharomyces cerevisiae • Buffer: 0.05M Malonate with 6.0M Urea, pH 5.2 ± 0.2 HSV-2 Recombinant Antigen, Glycoprotein G 2 • Represents unique sequences not present in HSV-1 • Fused with superoxide dismutase 1 (SOD) • Produced in Saccharomyces cerevisiae • Buffer: 50mM NaH 2 PO 4 , 160mM KCl, 5mM DTT, pH 7.0 ± 0.1 HSV-2 Antigen Glycoprotein G (gG) • Made synthetically > 99% pure (Analytical HPLC and Mass Spectrometry) • Contains the HSV-2 gG immunodominant regions • Immunoreactive with HSV positive sera • Buffer: 25 mM Tris-HCl, pH 8 HSV-2 Recombinant Antigen Glycoprotein G (gG) • Contains the gG-2 prepared by expressing the gene US4 from HSV-2 • Molecular Weight (Calculated): 35.8 kDa • Produced in E. coli , ~95% pure (12.5% SDS-PAGE) • Buffer: 20 mM PBS @ pH 7, 1 M NaCl

IgM Detection & Type specific for ELISA and CLIA Assays

VTI530

R01594

IgM Detection & Type Specific for ELISA

R01673

IgM Detection & Type Specific for ELISA, CLIA & LF Assays

C05014MA

MAb to HSV-1 Nucleocapsid protein (155kDa) • Reacts with HSV-1 Glycoprotein G 1 • Cross-reacts with HSV-2 nuclear protein

IFA Detection

C66150M

MAb to HSV-1 Glycoprotein G 1 • Reacts with HSV-1 Glycoprotein G 1

C01859M

MAb to HSV-2 Glycoprotein D (gD) • Reacts with HSV-2 Glycoprotein gD

ELISA & IFA Detection Assays

17

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Varicella Zoster Virus (VZV)

VZV is one of eight herpes viruses and commonly causes chickenpox in children, teens and young adults and herpes zoster (shingles) in adults. It is usually a mild disease that lasts a short time in healthy children. However, it can be severe in adults and may cause serious and even fatal complications in people of any age.

VZV infects the nerves causing a wide variety of symptoms and two clinically distinct forms of disease. Primary infection results in chickenpox and recurrent infections leads to herpes zoster (shingles). Symptoms of VZV are exhibited between 10 and 21 days after infection. The main symptom is a rash that turns into open lesions which crust over. It is spread through the airborne route primarily from the skin vesicles. The immunologic mechanism that controls latency of VZV is not well understood, however factors associated with recurrent disease include aging, immunosuppression, intrauterine exposure to VZV, and having had varicella at a young age (younger than 18 months). Infection with VZV is generally mild and self-limited, but it may be associated with complications, especially in adults or children under 1-year old. Secondary bacterial infections of skin lesions with Staphylococcus or Streptococcus are the most common cause (5% of cases) of hospitalization and invasive group A streptococci can lead to serious illness or death. Other complications include secondary pneumonia (bacterial or viral), aseptic meningitis, encephalitis, myocarditis and arthritis. A more rare complication is Reye’s syndrome and occurs almost exclusively in children who take aspirin during the acute illness. Routine vaccination against VZV is performed in the United States and Japan, however most countries still do not vaccinate. It is now available

VARICELLA VIRUS

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Source: MedicineNet, Inc.

as a quadrivalent measles-mumps-rubella- varicella (MMRV) vaccine and it is gaining wider acceptance globally. The VZV vaccine is on the World Health Organization’s (WHO) List of Essential Medicines, a list of the most important medications needed in a basic health system.

CONGENITAL VZV INFECTION

DIAGNOSIS The clinical presentations of VZV are very characteristic, however diagnosis is important for determining the immune status before prognostic and therapeutic monitoring. Several methods exist including polymerase chain reaction (PCR), direct immunofluorescent assay (DFA), viral isolation and serologic assays that detect VZV-specific antibodies. Recent infection is suggested by the detection of serum VZV-specific IgM antibodies, but this can be less reliable for herpes zoster where specific antibodies are already present. The National VZV Laboratory at the CDC has developed a reliable IgM capture assay. Other current commercials assays for determining VZV immune status include ELISAs, latex agglutination, indirect-immunofluorescence assay (IFA) and enzyme-linked fluorescent immunoassay (ELFAs). Primary maternal varicella infection in the first 20 weeks of gestation is associated with a variety of abnormalities in the newborn collectively known as congenital varicella syndrome. The range and severity of associated symptoms and physical findings may vary greatly from case to case depending upon when the maternal varicella zoster infection occurred during fetal development. In general, newborns with congential VZV have a low birth weight, distinctive skin abnormalities, and brain malformations.

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ToRCH & Childhood Diseases - Reagents for Assay Development

REAGENTS FOR SEROLOGY TESTING

7209

VZV Native Antigen • Prepared from infected (Ellen strain) HF cells

• Partially purified >10% viral protein • Buffer: 0.1M Glycine, pH 9.3-9.7

7201

VZV Native Antigen (PBS buffer) • Prepared from infected (Ellen strain) HF cells • Partially purified >10% viral protein • Buffer: PBS, pH 7.3-7.7 VZV Grade II Native Antigen (detergent free) • Prepared from infected (Ellen strain) HF cells • Protein concentration: 0.2-1.0mg/mL by OD260/280 • Buffer: 0.1M Glycine, pH 9.3-9.7

IgG Detection for EIA Assays

7740

C05100MA

MAb to VZV • Rotavirus group A specific antigen VP6 • Reacts with strains EDIM, SA-11, WA, and bovine NCDV strains

Paired Ag/Ab for IgM Capture Assays

VZV Native Antigen 7209

C05101MA

MAb to VZV (gpI & IV) • Reacts with precursor as well as with mature VZV glycoprotein I and glycoprotein IV

C05102MA

MAb to VZV (gpII) • Reacts with the carboxy region of VZV glycoprotein II (VZVgB)

C05104MA

MAb to VZV (gpIII) • Reacts with VZV glycoprotein III

C05105MA

MAb to VZV (175 kDa, Gene 62) • Reacts with VZV immediate early protein encoded by gene 62 MAb to VZV (gpIV) • Reacts with VZV glycoprotein IV (VZVgI) and to a lesser extent VZV glycoprotein I (VZVgE) by immunoprecipitation test • Reacts with both precursor and mature glycoprotein IV (VZVgI) MAb to VZV (Mixed) • A mixture of MAbs to VZX - includes CAT# C05100MA, C05101MA, C05102MA, C05104MA, C05105MA and C05107MA

IFA Detection

C05107MA

C05108MA

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Epstein-Barr Virus (EBV)

The Epstein–Barr virus (EBV), also called human herpesvirus 4 (HHV-4), is a virus of the herpes family and infects more than 95% of the world’s population. The most common manifestation of primary infection with this organism is acute infectious mononucleosis, a self-limited disease that frequently affects adolescents and young adults.

Primary EBV infections are typically asymptomatic and it is perhaps the most common reason for fever of unknown origin in young children. It does not occur in epidemics and has relatively low transmissibility, spreading mainly through bodily fluids (saliva). Approximately 90% of the US population is infected with EBV by the age of 25 and this infection rate is similar in other developed countries worldwide. EBV during pregnancy and transplacental transmission is rare. When infection with EBV occurs during adolescence or young adulthood, it causes infectious mononucleosis 35% to 50% of the time. EBV is the first human virus to be directly implicated in carcinogenesis. Infection is associated with particular forms of cancer, such as Hodgkin’s lymphoma, Burkitt’s lymphoma, nasopharyngeal carcinoma, and conditions associated with AIDS including hairy leukoplakia and central nervous system lymphomas. In particular in Africa, the virus is associated with endemic Burkitt lymphoma in DIAGNOSIS Diagnosing an EBV infection can be challenging since the symptoms are similar to other illnesses. However, different proteins are expressed during the various stages the EBV life cycle and the detection of these antigens can help distinguish whether an infection is a primary acute, convalescent, latent, or reactivation infection. Diagnostic methods for EBV include IFA, ELISA, blot techniques, IgG avidity, PCR and virus isolation. About 90% of adults have antibodies that show that they have a current or past EBV infection. In most cases, the antibody response occurs rapidly during primary EBV infection. Tests are available for detecting antibodies to the following EBV-associated antigens: Viral capsid antigen (VCA) •  Elevated anti-VCA IgM indicates acute infection: appears early in infection and usually disappears within 4 - 6 weeks

EBV INFECTION KINETICS

Source: katkars.com

the setting of co-infection with Plasmodium falciparum. Specifically, it has been found that a malaria infection can impair the T-cell response to EBV and directly contribute to tumor pathogenesis.

• Elevated anti-VCA IgG indicates prior infection: appears in the acute phase, peaks 2-4 weeks after onset, declines slightly, then persists for the rest of a person’s life • Common VCA proteins are gp125 and p19 Early antigen (EA) • Anti-EA IgG appears in the acute phase of illness and generally falls to undetectable levels after 3-6 months • In many people, detection of antibody to EA is a sign of an active infection • 20% of healthy people may have antibodies against EA for several years EBV nuclear antigen (EBNA) • Antibody to EBNA is not seen in the acute phase of EBV infection but slowly appears 2-4 months after onset of symptoms • Persists for the rest of a person’s life

20

ToRCH & Childhood Diseases - Reagents for Assay Development

REAGENTS FOR SEROLOGY TESTING

7420

EBV Native Antigen • Prepared from a detergent extraction of P 3 HR-1 or P 3 H 3 cells that have been induced by TPA and sodium butyrate • Partially purified >10% viral protein • Buffer: Carbonate, pH 9.5±0.2 EBV EBNA-1 Recombinant Antigen (full length) • Represents full length EBNA-1 and contains a His-tag at the N-terminus • Produced in E. coli • Buffer: 6M Urea, 20mM Phosphate, pH 8.0, 1M Sodium Chloride, 0.1% Polyoxyethylene(10)Tridecylether EBV p138 (EA-D) Recombinant Antigen • Represents the complete ORF of the BALF2 gene (Early Antigen D) • Contains a His-tag at the N-terminus • Produced in E. coli • 6M Urea, 20mM Phosphate, pH 8.0, 1M Sodium Chloride, 0.1% Polyoxyethylene(10)Tridecylether EBV VCA (gp125) Antigen • Purified antigen concentrate from P 3 H 3 cells • Concentration: ~ 30 μg/mL (Non-Interfering Protein Assay) • Buffer: 20 mM Tris-HCl, pH 7.4 containing 3M Magnesium Chloride

IgG Detection for ELISA and WB Assays

R01522

8202

IgM Detection for ELISA, WB, Lateral Flow and CLIA Assays

R01525

C66405M

MAb to EBV (gp125) • Reactive against EBV VCA gp125

C65023M

MAb to EBV (VCA) • Recognizes EBV, specific for VCA MAb to EBV (EBNA-1) • Specific for envelope glycoprotein complex 220/350

IFA Detection

C65221M

• EBV glycoprotein gp220/350 is the major glycoprotein associated with the EBV envelope (the 220 kd protein is the result of RNA splicing)

EBV VCA p23 Recombinant Antigen • Represents the ORF of the BLRF2 gene (a.a. 2-162) coding for the EBV tegument protein BLRF2 (23kDa) • Produced in E. coli (> 95% pure SDS-PAGE), lyophilized • Lyophilized from 20 mM Phosphate Buffer, pH 8, 1 M Sodium Chloride EBV VCA p18 Recombinant Antigen • Represents the truncated form of the viral capsid protein VP 26 (EBV gene BFRF3)

R01570

IgM Detection for ELISA and WB Assays

R01571

• Produced in E. coli ( ≥ 98% pure SDS-PAGE) • Buffer: 20 mM PBS @ pH 7 and 1 M NaCl

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Mumps

Mumps is an enveloped single-strand RNA virus of the Rubulavirus genus that causes painful swelling of the salivary glands. It is highly contagious and predominantly affects children.

The mumps virus resides in the mucus of the nose and throat of an infected person and it is mainly spread through coughing and sneezing. Symptoms may not appear for 12-25 days after transmission, however an infected person is contagious from 3 days prior to the onset of symptoms to 9 days after. In general, only supportive care is needed to resolve a mumps infection but occasionally it causes serious complications including meningitis, encephalitis, deafness and orchitis (inflammation of the testicles in males). Before the routine vaccination program was introduced in the United States (in 1967) and other countries, mumps was a common illness in infants, children and young adults. However, due to vaccination the disease is becoming rare. DIAGNOSIS

COUNTRIES USING MUMPS VACCINE IN NATIONAL IMMUNIZATION SCHEDULE, 2012

No (74 countries or 38%) Yes (120 countries or 62%) Not available Not applicable

Standard assays that detect mumps include viral detection methods, RT-PCR and serologic assays in both EIA and IFA formats. Specifically, assays that detect both IgM and IgG antibodies work well for diagnosing mumps infection in immunologically naïve individuals. However, the IgM response and viral shedding that occurs in persons who have been previously vaccinated or naturally infected are moderate in duration and intensity, making detection difficult. Recent research has shown that the enzyme-linked immunospot (ELISPOT) assay could be used as a more reliable diagnostic. This immunoassay method is based on the sandwich ELISA technique and is highly sensitive and the enables detection of activated mumps-specific, antibody-secreting B cells in whole blood.

REAGENTS FOR SEROLOGY TESTING

8099

Mumps Native Antigen • Prepared from a glycine extraction of infected (Enders Strain) LLC-MK2 cells • >10% viral protein, partially purified to reduce host cell components • Protein concentration: 0.2-1.0mg/mL by OD 260/280nm • Buffer: 0.1M Glycine, pH 9.3–9.7 Mumps Grade III Native Antigen • Prepared from infected (Enders Strain) LLC-MK2 cells • Purified by ion exchange chromatography to remove host cell protein • Buffer: 0.1 Na 2 CO 3 , 0.1M NaCl, pH 7.9-8.3

IgG Detection for EIA Assays

EV9529

IgM Detection for EIA Assays

C01719M C01720M C01721M

MAb to Mumps Virus Nucleoprotein • Reacts with Mumps Virus Nucleoprotein

ELISA & IFA Detection Assays

22

ToRCH & Childhood Diseases - Reagents for Assay Development

Rubeola (Measles)

Rubeola (also known as measles) is a highly contagious viral infection of the respiratory system, immune system, and skin. Rubeola is caused by a paramyxovirus of the genus Morbillivirus .

RECENT INCREASE IN MEASLES – USA CASES BY YEAR

Complication with measles is common and can be fatal. It is a leading cause of vaccine-preventable childhood mortality. The classic signs and symptoms of measles include a high fever, coughing, conjunctivitis and a characteristic rash. Symptoms usually develop 7–14 days after exposure and 3 out of 10 people who get measles will develop one or more complications including pneumonia, encephalitis and death. There is no specific treatment for measles although in developed countries, children are immunized against measles at 12 months old as part of a three-part Measles, Mumps and Rubella vaccine. A recent increase in measles worldwide has been driven by unvaccinated people spreading the disease into countries where it was once declared eliminated. Measles is extremely infectious and among unimmunized people exposed to the virus, over 90% will contract the disease. DIAGNOSIS

Measles-specific IgM serology is the standard test for rapid laboratory diagnosis of measles. IgM testing using ELISA indirect capture methods or measles IgM capture are almost exclusively used. These assays require a pretreatment step to remove IgG antibodies and rheumatoid factor to ensure optimal performance. However, in regions where endemic measles virus has been eliminated, additional diagnostic assays are used to confirm measles cases irrespective of vaccination status. IgG avidity testing has proved useful in cases that require additional methods, such as suspected false negatives or a false positives.

REAGENTS FOR SEROLOGY TESTING

7604

Rubeola Native Antigen • Prepared from a glycine extract of infected (Edmonston strain) Vero cells • >10% viral protein, partially purified to reduce host cell components and contains predominantly nucleocapsid antigens • Buffer: 0.1M Glycine, pH 9.3–9.7 Rubeola Virus Nucleoprotein Recombinant Antigen • Produced in Sf9 insect cells, sequence derived from the Edmonston strain • Protein Concentration: 0.5 ± 0.1mg/mL by BCA • Buffer: 50mM Hepes, pH 7.5 ± 0.1

IgG Detection for EIA Assays

EV9298

IgM Detection for EIA Assays

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