MRSA (MREJ, mecA, mecC, vanA, MSSA) direct PCR Kit CE-IVDR

The MRSA direct PCR is a qualitative assay for in vitro examination of bacterial DNA in nasal/throat swabs, swabs from skin/wounds or culture from patients to provide information to aid to diagnose symptomatic or asymptomatic patients under suspicion of infection with methicillin-resistant Staphylococcus aureus (MRSA) and/or vancomycin-resistant Staphylococcus aureus (VRSA) as well as Methicillin-susceptible Staphylococcus aureus (MSSA).
The IVD medical device detects the DNA of the aforementioned pathogens by qualitative measurements based on qPCR and is intended for use in medical laboratories or health institutions by laboratory personnel specifically trained in qPCR and in vitro diagnostic techniques. It has to be used in combination with conventional qPCR cyclers for detection and analysis.

Testing Principle
The MRSA direct PCR test is a multiplex qPCR test for the detection of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Staphylococcus aureus (VRSA) as well as Methicillin-susceptible Staphylococcus aureus (MSSA).
The assay can be performed either on purified nucleic acids obtained through a prior extraction procedure (cf.cmanual) or directly on the sample material without prior isolation and purification (cf. manual).
For workflows including nucleic acid extraction, a separate Internal Control (IC) is added to each sample prior to RNA extraction and serves as control for nucleic acid isolation from the biological specimen as well as for amplification.
For direct testing without isolation and purification, a separate solution (Solution B) is provided containing an alternative Internal Control that functions as an amplification control, only.
The nucleic acid eluate or direct sample material is added to the ready-to-use reaction solution that contains all reagents necessary for qPCR. The analysis can be performed on various qPCR cyclers.
Possible sample material from different sources (nasal/throat swabs, swabs from skin/wounds or culture) include:- samples provided as dry swabs
- samples provided as Amies Agar gel swabs
- samples provided in Copan eSwabsTM
- samples provided in Copan UTM-RT System (UTM-RT)
- samples provided in Yocon VTM/UTM
- samples provided in NaCl or PBS solution
- samples provided in Amies solution
- samples provided in or in IsoFlow Steath Fluid (Beckman Coulter)
- Tryptic Soy Broth (TSB)
The MRSA direct PCR test contains probes specific for the following targets: (1) MREJ, (2) mecA, (3) mecC, and (4) Internal Control. The probes are each labelled with fluorescent reporter dyes and a second dye that serves as a quencher and suppresses the fluorescence signals of the intact probes. The analysis is performed by determining Ct (cycle threshold) values. The Ct value describes the cycle in which the signal rises above a certain threshold for the first time. The more target copies (here: bacterial DNA) are present in the sample, the lower the Ct value. The Internal Control (IC) allows the detection of PCR inhibition. The buffer contains additives for bacterial lysis and enhancement of PCR efficiency. Patient samples can be analysed directly without prior DNA extraction.

Features
- Analyte(s): orfX, MecA, MecC, spa for MRSA; vanA , spa for VRSA; spa for MSSA
- Sample origin: Human
- Sample type: nasal swab
- Assay type: qPCR (Realtime PCR)
- qPCR cycler detection channels: Cyan500/FAM/HEX/Red610/Cy5 for MRSA, VRSA, MSSA with internal control Cyan500 (catalog nr. FBC131-Ax); FAM/HEX/Red610/Cy5/Cy5.5 for MRSA, VRSA, MSSA with internal control Cy5.5 (catalog nr. FBC131-Bx); FAM/HEX/Red610/Cy5 for MRSA; MSSA with internal control HEX (catalog nr. FBC131-Cx)
- Quantity: 96 reactions/kit
- Certification: EU-IVDR

Target information:
- MRSA (orfX) FAM, VRSA (vanA) HEX, MRSA (mecA/MecC) Red 610, S. aureus (MSSA) (spa) Cy5, Internal Control (Artifical NA) CY500 (FBC131-Ax)
- MRSA (orfX) FAM, VRSA (vanA) HEX, MRSA (mecA/MecC) Red 610, S. aureus (MSSA) (spa) Cy5, Internal Control (Artifical NA) CY5.5 (FBC131-Bx)
- MRSA (orfX) FAM, MRSA (mecA/MecC) Red 610, S. aureus (MSSA) (spa) Cy5, Internal Control (Artifical NA) HEX(FBC131-Cx)
The MRSA direct PCR test was validated with the Roche LightCycler® 480 II and with the BioRad CFX Opus 96™.
In general, Lab direct qPCR tests are compatible with other qPCR cyclers (e.g., MIC Cycler, AJ qTOWER, LightCycler® 96 or 480 I).