Recent studies have documented an increase in the incidence of antifungal resistance in newly emerging species closely related to C. albicans, and the coexistence of genotypic variants. Hence, an application of PCR-based molecular typing is crucial in identifying these fungi. Our study used molecular methods to characterize the latest genotypic subgroups of C. albicans and analysed if there was a relationship between the genotypes and the antifungal resistance pattern. The study was conducted in JSS Hospital, Mysuru, Karnataka between July 2018 and December 2020. A total of 1427 Candida species were isolated from clinical samples. Candida albicans were isolated and confirmed using Germ tube test, ID VITEK 2 and PCR (ITS primer). DNA extraction was done using the Hi-Media Yeast DNA Extraction Kit. The amplified products were analysed using Agarose gel electrophoresis (2%). Among 1427 Candida species, 282 were Candida albicans. The following resistance was exhibited to major antifungals – Caspofungin (3.5%), Amphotericin B (1.4%), flucytosine (2.8%) Fluconazole (6%) Micafungin (2.8%) Voriconazole (3.1%) and all were sensitive to miconazole. ABC genotyping showed Genotype A (450 bp) predominant (87.58%) followed by genotype B (840bp) (9.92 %) and genotype C (450bp and 840 bp) (0.2%). Genotype D and E were not observed. Our study showed the growing antifungal resistance in clinical isolates. Genotype A was predominant in South Karnataka region followed by Genotype B and C. There was no correlation between genotyping and antifungal resistance. However, a study with greater number of samples from diverse geographical locations may give more insight.
COVID-19 detection via lateral flow antigen assays (LFA) are rapid and economically acquiescent to infrastructure facile healthcare settings. Early, prompt identification of cases to facilitate patient isolation and supportive management is the essence of rapid diagnostic tests. Given the backdrop of post COVID-19 pandemic-molecular testing still remains a costly affair. Additionally, molecular assays are incapable of distinguishing remnant RNA from replication competent viruses. In this scenario, we explore the diagnostic consonance of SARS-CoV-2 LFAs with RT-PCR cycle threshold, in a likelihood that it could be used as a surrogate marker for infection transmissibility. Rapid COVID-19 LFA results were compared with Real-time PCR for detection of SARS-CoV-2 in nasopharyngeal swabs. Two hundred rapid antigen positive nasopharyngeal swabs obtained from COVID-19 suspects/contacts/preoperative/screening patients were subjected to RT-PCR to study the correlation with cycle threshold (CT) values obtained for all the antigen positive cases. 200 Rapid COVID-19 LFA positive samples were analyzed in the present study. Amidst the LFA positive samples included in the study 187 (93.5%) were found to have concordant results when subjected to the gold standard Real-time PCR. Discordant results were documented in 13 (6.5%) COVID-19 LFA positive samples which were found to be negative by RT-PCR. The average Cycle threshold values were found to be 23.75 for E gene, 25.36 for N gene and 24.07 for RdRp gene. The average PCR Cycle threshold of LFA positive cases remained significantly undeterred (p<0.5) throughout the time period of the study stipulating the undaunted viral load across the different waves of the pandemic. Maximum association of LFA positivity with symptom-manifestation was seen during the 1st wave of COVID-19 (September-December 2020 in India). The association of symptoms with LFA test positivity reduced to a significant extent during the 3rd wave of the pandemic in January 2022 (p<0.5) indicating the reduced clinical severity but not infectivity of the SARS-CoV-2 infection during the 3rd wave of the pandemic. Lateral flow assay based diagnostic tests are technically & economically convenient modalities with significant interest concordance in comparison with RT-PCR. Definitive advantage in terms of achieving quick patient triage and thereby patient management can be achieved with the use of these tests.
Staphylococcus aureus strains that are mecA and PBP2a positive but phenotypically susceptible to oxacillin are becoming more and more abundant, according to research from all around the world. The oxacillin susceptibility of Staphylococcus aureus (OS-MRSA) contributes to consequent treatment-failure due to misidentification by conventional susceptibility tests. Therefore, the objective of the current study was to ascertain the prevalence of OSMRSA in a tertiary care facility located in Mysore, South India. 395 MRSA isolates collected from diverse clinical samples were included in this lab-based prospective investigation. These isolates were tested using an oxacillin 1μg disc phenotypically by standard disc diffusion test, and simultaneously MIC to Oxacillin was determined from Vitek2 systems. Additionally, MRSA specific mecA gene detection was applied to these isolates in order to confirm their MRSA status genotypically. PCR findings demonstrate that 65% of the isolates were MRSA. The vitek2 system detected 4.06% OS-MRSA isolates with an oxacillin MIC of ≤2µg/ml. The disc diffusion method identified a total of 13.75% isolates as oxacillin sensitive and 10% isolates were oxacillin intermediately sensitive. Oxacillin sensitivity was shown for 1.87% of the mecA-positive MRSA isolates using the VITEK2 and disc diffusion techniques. This analysis found isolates with lower oxacillin MICs but relatively reduced OS-MRSA incidence. Using an oxacillin disc for routine laboratory MRSA detection might occasionally produce false negative results, which can result in improper antibiotic administration and treatment failure. In order to distinguish OS-MRSA from MRSA, it is crucial to combine phenotypic and genotypic techniques.
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