Lyo-Ready TM Direct qPCR Urine Mixes Specimen-specific TM mixes designed for fast, direct DNA or RNA amplification from urine specimens
Meridian’s Lyo-Ready Direct DNA qPCR Urine and RNA/DNA qPCR Urine mixes are ready-to-use, 4x concentrated, fully optimized mixes designed for rapid, sensitive amplification of RNA or DNA from crudely processed urine specimens. Urine specimens have grown in popularity for testing sexually transmitted disease (STDs) as they are less invasive than a cervical or urethra swab and the easy sample collection encourages routine screening. More than 30 bacteria, viruses and parasites are known to be transmitted through sexual contact, however only 8 pathogens (syphilis, gonorrhoea, chlamydia, trichomoniasis, hepatitis B (HBV), herpes simplex virus (HSV), HIV and human papillomavirus (HPV)) are responsible for the majority of infections. The prevalence of STDs has increased over the past several years, and according to the 2019 STD Surveillance Report released by the Centres for Disease Control (CDC), there was a 30% increase in reportable STDs between 2015 and 2019. The World Health Organization also estimates that 1 million sexually transmitted infections are acquired globally each year and most of them are asymptomatic. Studies have shown that the stigma attached to STDs, limited access to screening tests and the lack of any disease symptoms, all undermine STD testing and treatment, contributing to the further spread of disease. At home testing is a growing trend that alleviates many of these issues and several testing kits already exist for a variety of STDs including gonorrhea, chlamydia, syphilis and HIV. However most of these tests still require a swab or blood prick which can be challenging for self-collection. Urine is an alternative specimen type that enables easy self-sampling and studies have shown that it is just as effective for screening many STDs, in particular bacterial STDs such as chlamydia and gonorrhea. Lyo-Ready TM Direct DNA qPCR and Direct RNA/DNA qPCR Urine mixes are designed to deliver fast and sensitive amplification of DNA or RNA using direct detection methods. Patient urine sample can be used on the dried assay and do not require nucleic acid
purification. The mixes also contain excipients and an optimized buffer system that is compatible with lyophilization to create ambient-temperature stable assays which have more than two year shelf-life.
• Ideal for developing screening assays for sexually transmitted diseases (STDs) and urinary tract infections (UTIs) • Very sensitive detection of low copy number bacteria, viruses and cell-free nucleic acids from crude urine samples, even using extraction-free protocols • Quickly and accurately detect slow-growing and difficult-to-culture microorganisms, removing ambiguous results • Can be kept as a liquid mix or lyophilized to create ambient-temperature stable assays with an extended shelf-life
CAT NO. VOLUME REACTIONS
5 mL 1,000 Rxns 50 mL 10,000 Rxns 5 mL 1,000 Rxns 50 mL 10,000 Rxns
Lyo-Ready Direct DNA qPCR Urine
Lyo-Ready Direct RNA/DNA qPCR Urine
Superior reaction efficiency using crude urine samples in multiplex reactions
Lyo-Ready™ Direct DNA qPCR Urine (MDX152)
MDX152 | Quantabio ToughMix | Roche KAPA | ThermoFisher TaqPath ™
Four sexually transmitted pathogens ( M. genitalium, T. pallidum (Syphilis), C. trachomatis and N. gonorrhoeae ) were amplified in a quadruplex reaction in the presence of 20% human urine using Lyo-Ready ™ Direct DNA qPCR Urine in a lyophilized format ( red ) and compared to Quantabio ToughMix PCR Master Mix ( grey ), Roche KAPA Probe Force qPCR kit ( green ) and ThermoFisher TaqPath ™ ProAmp ™ Multiplex Master Mix ( black ) in a liquid format. The results demonstrate that Lyo-Ready ™ Direct DNA qPCR Urine has higher multiplexing capabilities and better reproducibility than other supplier qPCR mixes.
Lyo-Ready™ Direct RNA/DNA qPCR Urine (MDX153)
MDX153 | ThermoFisher TaqPath ™ | Quantabio ToughMix | BioRad Reliance 1-Step
Three viral RNA (Dengue, Zika and Chikungunya) and one DNA (Malaria) mosquito-borne infectious disease targets were amplified in a quadruplex reaction in the presence of 20% human urine using Lyo-Ready ™ Direct RNA/DNA qPCR Urine in a lyophilized format ( red ) and compared to ThermoFisher TaqPath ™ 1-Step Multiplex Master Mix ( black ), Quantabio UltraPlex 1-Step ToughMix ™ ( grey ) and BioRad Reliance 1-Step Multiplex RT-qPCR Supermix ™ ( pink ) in liquid formats. The results illustrate that dry Lyo-Ready ™ Direct RNA/DNA qPCR Urine has higher multiplexing capacity and reproducibility than the liquid competitor mixes.
High amplification sensitivity with low copy templates using either liquid and dried-down assay formats
Lyophilized | Liquid
Activity of A) Lyo-Ready ™ Direct DNA qPCR Urine in lyophilized ( red ) and liquid ( blue ) format was compared in the ability to amplify a 10-fold serial dilution of synthetic DNA (10,000, 1,000, 100 and 10 copies respectively) and B) Activity of Lyo-Ready ™ Direct RNA/DNA qPCR Urine in lyophilized ( red ) and wet ( blue ) format was compared in the ability to amplify a 10-fold synthetic RNA target (10,000, 1,000, 100 and 10 copies respectively) in the presence of 10% pooled human urine. The results illustrate that the dried mix retains the ability to efficiently amplified to the same level as the wet mix with the same level of sensitivity and reproducibility for both.
Fast and more sensitive detection of cancer markers in urine samples
Three tumor-related mRNA markers (CDC2 kinase, IGFBP5 and MDK) were amplified from total RNA (from a bladder cancer patient) in a triplex reaction using lyophilized Lyo-Ready ™ Direct RNA/DNA qPCR Urine ( red ) or liquid BioRad Reliance One-Step Multiplex Supermix ™ ( pink ) in the presence of 10% Human Urine. The results illustrate that Lyo-Ready ™ Direct RNA/DNA qPCR Urine is able to detect cancer related RNA markers from urine faster and with higher sensitivity compared to the other mix.
MDX153 | BioRad Reliance 1-Step
Compatible with very fast cycling conditions to deliver very fast results
A/ Lyo-Ready™ Direct DNA qPCR Urine (MDX152)
A) Targets from Neisseria gonorrhoeae , Chlamydia trachomatis and group B streptococcus (GBS) were amplified in a triplex reaction in the presence of 25% human urine under standard cycling conditions: 40 cycles of 10 sec @ 95°C and 30 sec @ 60°C and fast cycling conditions: 3 min at 95°C followed by 40 cycles of 1 sec @ 95°C and 1 sec @ 60°C. B) Three mammalian RNA targets were amplified in a triplex reaction in the presence of 5% human urine under standard cycling conditions: 10 min reverse transcription step followed by 40 cycles of 10 sec @ 95°C and 30 sec @ 60°C and fast cycling conditions: 5 min reverse transcription step followed by 40 cycles of 1 sec @ 95°C and 1 sec @ 60°C. All reactions were carried out in the Quantstudio ™ 7 Flex Real-Time PCR System. The results illustrate the ability of Lyo-Ready Direct Urine mixes to deliver the fastest possible results, without compromising the sensitivity and reproducibility of an assay.
B/ Lyo-Ready™ Direct RNA/DNA qPCR Urine (MDX153)
Extended shelf-life of greater than 12 months at ambient temperature
Lyophilized | Liquid
Lyo-Ready ™ Direct DNA qPCR Urine and Lyo-Ready ™ Direct RNA/DNA qPCR Urine were lyophilized, and the stability tested in accelerated stability studies. The lyophilized mixes ( red ) were incubated at 37°C for 1 month and tested against the fresh liquid mixes ( blue ) in a triplex qPCR assay with 10% human urine (MDX152) and a triplex RT-qPCR assay with 5% human urine (MDX153). Results suggest that the lyophilized mix is active following accelerated stability tests with projected at least 12 months stability at ambient temperature.
Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Phone: +1 901-382-8716 Fax: +1 901-333-8223
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