Applied Biosystems™ MycoSEQ™ Plus Mycoplasma Detection Kits
Introduction
The Applied Biosystems™ MycoSEQ™ Plus Mycoplasma Detection Kit is a TaqMan™-based quantitative PCR (qPCR) kit for the detection of mycoplasma in complex bioproduction samples. Designed and tested using criteria often applied toward rapid mycoplasma detection in biotherapeutic manufacturing cell culture lot release, the MycoSEQ Plus Kit enables results that meet or exceed guidance on sensitivity as described in the European Pharmacopoeia (E.P. 2.6.7, 2007), the US Pharmacopoeia (US63), and the Japanese Pharmacopoeia. When used with a suitable sample preparation method, the MycoSEQ Plus Kit can detect less than 10 CFU/mL or the genomic equivalent of 10 GC/mL. This multiplexed assay enables predicted simultaneous screening of over 200 species of mycoplasma target, discriminatory detection of an assay-specific control template, and internal PCR control with no cross-reactivity to closely-related bacterial species.
Features of the Detection Kit
Notable aspects of the MycoSEQ Plus Mycoplasma Detection Kit include:
- Results provided in less about 5 hours
- A unique discriminatory positive control to differentiate a true positive result from a potential cross-contamination positive call
- Protocols for various methods
- Live mycoplasma is not needed. The Kit does not contain any live mycoplasma and does not require the use of live mycoplasma for validation
- Can be used with gDNA, inactivated mycoplasma or live mycoplasma
- May be used as an alternative to the standard 28-day culture test
- May be used in conjunction with culture-based methods by providing preliminary results while waiting for results from the 28-day test
Complementary Sample Prep
Applied Biosystems™ PrepSEQ™ Nucleic Acid Extraction Kits offer a robust and easy-to-follow workflow to overcome complex matrix challenges and ensure the required sensitivity. MycoSEQ Plus Mycoplasma Detection Kits should be used with the PrepSEQ 1-2-3 Nucleic Acid Extraction Kit (Cat. No. 18775505) for manual extractions, or with the PrepSEQ Express Nucleic Acid Extraction Kit (Cat. No. 18755455) for automated extraction using the Applied Biosystems™ AutoMate Express™ Nucleic Acid Extraction System (Cat. No. 30055377). Also available but not included with the detection kit are two MycoSEQ gDNA Kits (Cat. Nos. 18705395 and 18725495), which contain purified genomic DNA from various mycoplasma species and may be used as positive test materials during method development.
An Integrated Workflow Solution
The MycoSEQ Plus Mycoplasma Detection Kit is part of an integrated workflow for adventitious agent, impurity, and contaminant testing during biopharmaceutical manufacturing. The entire workflow, including PrepSEQ Sample Preparation Kits which offer manual or automated sample preparation approaches, together with qPCR analysis, can provide results in under 5 hours. (Please note complex matrices may require additional upfront processing steps, as described in various available protocols.)
MycoSEQ Plus Kit performance has been tested and confirmed on the Applied Biosystems™ 7500 Fast Real-Time PCR System and the Applied Biosystems™ QuantStudio 5 Real-Time PCR System. Data analysis using Applied Biosystems™ AccuSEQ™ Real-Time PCR Detection Software provides accurate quantitation and security, audit, and e-signature capabilities to help enable 21 CFR Pt 11 compliance.
