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Introduction: mecC Methicillin Resistant Staphylococcus aureus (MRSA) after its initial recovery in 2007 has been reported with varying frequency from different parts of the world. These isolates assume importance due to the fact that with routine testing platforms available for the detection of MRSA they can be misidentified as being methicillin sensitive which can adversely affect the treatment and outcome of infections due to MRSA harbouring the mecC gene. Aim: Evaluation of mecC gene carrying MRSA in clinical isolates. Materials and Methods: This retrospective study was conducted in the Department of Microbiology, Sher-i-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India for a period of three months (May-July 2020). A total of 102 laboratory confirmed isolates of MRSA (based on biochemical tests and cefoxitin disc diffusion results) were subjected to screening for the presence of mecA and mecC gene by Polymerase Chain Reaction (PCR). Deoxyribonucleic Acid (DNA) was extracted using an in house extraction method following which mecA and mecC were amplified in a total reaction volume of 25 μL using 2x PCR master mix, 5 μL of template and 1 μL (0.4 μM final concentration) each of reverse and forward primers specific for the above mentioned genes. Statistical analysis was done using Statistical Package for the Social Sciences (SPSS) software v24.0. Results: All the isolates were confirmed as being methicillin resistant with 96.1% isolates carrying the mecA gene and 3.9% carrying the mecC gene. mecC MRSA were recovered from pus, swab and endotracheal tip in middle aged men. One of the patient from whose sample mecC MRSA was recovered was suffering from hypertension, diabetes and renal faliure. MRSA exhibited high resistance to all the antimicrobial agents tested however all of them were sensitive to vancomycin and linezolid. Conclusion: The presence of mecC gene in clinical isolates of MRSA is a cause of concern and calls for an extensive and continuous surveillance of such isolates as they can in future be implicated in causing severe human infections. [ABSTRACT FROM AUTHOR]

The mean Ct-values for all three genes (cumulative Ct-values) of samples collected in NS was compared to samples collected in VTM. Sir, The current pandemic of coronavirus disease-2019 (COVID-19) across the globe has exhausted the healthcare facilities including the diagnostic and sample collection centres across the world. A study from India has reported that NS gargle lavage may be a viable alternative to swabs for the collection of samples for COVID-19 diagnosis[[9]]. [Extracted from the article]

Background & Objectives: Public health and diagnostic laboratories are facing huge sample loads for COVID-19 diagnosis by real-time reverse transcription-polymerase chain reaction (RT-PCR). High sensitivity of optimized real-time RT-PCR assays makes pooled testing a potentially efficient strategy for resource utilization when positivity rates for particular regions or groups of individuals are low. We report here a comparative analysis of pooled testing for 5- and 10-sample pools by real-time RT-PCR across 10 COVID-19 testing laboratories in India., Methods: Ten virus research and diagnostic laboratories (VRDLs) testing for COVID-19 by real-time RT-PCR participated in this evaluation. At each laboratory, 100 nasopharyngeal swab samples including 10 positive samples were used to create 5- and 10-sample pools with one positive sample in each pool. RNA extraction and real-time RT-PCR for SARS-CoV-2-specific E gene target were performed for individual positive samples as well as pooled samples. Concordance between individual sample testing and testing in the 5- or 10-sample pools was calculated, and the variation across sites and by sample cycle threshold (C t ) values was analyzed., Results: A total of 110 each of 5- and 10-sample pools were evaluated. Concordance between the 5-sample pool and individual sample testing was 100 per cent in the C t value ≤30 cycles and 95.5 per cent for C t values ≤33 cycles. Overall concordance between the 5-sample pooled and individual sample testing was 88 per cent while that between 10-sample pool and individual sample testing was 66 per cent. Although the concordance rates for both the 5- and 10-sample pooled testing varied across laboratories, yet for samples with C t values ≤33 cycles, the concordance was ≥90 per cent across all laboratories for the 5-sample pools., Interpretation & Conclusions: Results from this multi-site assessment suggest that pooling five samples for SARS-CoV-2 detection by real-time RT-PCR may be an acceptable strategy without much loss of sensitivity even for low viral loads, while with 10-sample pools, there may be considerably higher numbers of false negatives. However, testing laboratories should perform validations with the specific RNA extraction and RT-PCR kits in use at their centres before initiating pooled testing., Competing Interests: None