Molecular epidemiology of carbapenem-resistant Enterobacteriaceae isolated from patients in COVID-19 wards and ICUs in a Bulgarian University Hospital.

Many studies report an increase in antimicrobial resistance of Gram - negative bacteria during the COVID-19 pandemic. Our aim was to evaluate the epidemiological relationship between carbapenem-resistant (CR) Enterobacteriaceae isolates from patients in COVID-19 wards and to investigate the main mechanisms of carbapenem resistance in these isolates during the period April 2020-July 2021. A total of 45 isolates were studied: Klebsiella pneumoniae (n = 37), Klebsiella oxytoca (n = 2), Enterobacter cloacae complex (n = 4) and Escherichia coli (n = 2). Multiplex PCR was used for detection of genes encoding carbapenemases from different classes (blaKPC, blaIMP, blaVIM, blaNDM, blaOXA-48). For epidemiological typing and analysis, ERIC PCR was performed. Two clinical isolates of E. cloacae, previously identified as representatives of two dominant hospital clones from the period 2014-2017, were included in the study for comparison. In the CR K. pneumoniae group, 23 (62.2%) carried blaKPC, 13 (35.1%) blaNDM, 10 (27.0%) blaVIM, and 9 (24.3%) were positive for both blaKPC and blaVIM. The blaKPC was identified also in the two isolates of K. oxytoca and blaVIM in all E. cloacae complex isolates. The two CR isolates of E. coli possessed blaKPC and blaOXA-48 genes. Epidemiological typing identified 18 ERIC profiles among K. pneumoniae, some presented as clusters of identical and/or closely related isolates. The carbapenem resistance in the studied collection of isolates is mediated mainly by blaKPC. During the COVID-19 pandemic intrahospital dissemination of CR K. pneumoniae, producing carbapenemases of different molecular classes, as well as continuing circulation of dominant hospital clones of multidrug-resistant E. cloacae complex was documented.

Dealing with inconclusive SARS-CoV-2 PCR samples-Our experience.

PURPOSE: Early confirmation of SARS-CoV-2 is a key point in the timely management of infected patients and contact persons. Routine diagnostics of COVID-19 cases relies on RT-PCR detection of two or three unique sequences of the virus. A serious problem for the laboratories is how to interpret inconclusive samples which are positive for only one of the SARS-CoV-2 specific genes. MATERIALS AND METHODS: A total of 16364 naso-oropharyngeal swabs were collected and tested with SARS-CoV-2 Real-TM kit (Sacace Biotechnologies, Italy) between May and September 2020. We retrospectively analyzed their amplification plots to determine the number of inconclusive samples. We also reviewed the medical records to summarize the patient's COVID-19 testing history and basic demographic characteristics. RESULTS: We obtained 136 (0.8%) inconclusive samples with amplification signal only for the N-gene. Thirty-nine of the samples were excluded from further analysis as no additional data were available for them. Of the rest of the samples, the majority- 48% (95% CI 38-59%) had a previous history of SARS-CoV-2 positivity, 14% (95% CI 8-23%)-a subsequent history of positivity and 37% (95% CI 28-48%) were considered as false positive. CONCLUSION: A substantial proportion of the inconclusive results should be considered as positive samples at the beginning or the end of the infection. However, the number of false-positive results is also significant and each patient's result should be analyzed separately following the clinical symptoms and epidemiological data.

Dealing with inconclusive SARS-CoV-2 PCR samples—Our experience

Purpose Early confirmation of SARS-CoV-2 is a key point in the timely management of infected patients and contact persons. Routine diagnostics of COVID-19 cases relies on RT-PCR detection of two or three unique sequences of the virus. A serious problem for the laboratories is how to interpret inconclusive samples which are positive for only one of the SARS-CoV-2 specific genes. Materials and methods A total of 16364 naso-oropharyngeal swabs were collected and tested with SARS-CoV-2 Real-TM kit (Sacace Biotechnologies, Italy) between May and September 2020. We retrospectively analyzed their amplification plots to determine the number of inconclusive samples. We also reviewed the medical records to summarize the patient’s COVID-19 testing history and basic demographic characteristics. Results We obtained 136 (0.8%) inconclusive samples with amplification signal only for the N-gene. Thirty-nine of the samples were excluded from further analysis as no additional data were available for them. Of the rest of the samples, the majority– 48% (95% CI 38–59%) had a previous history of SARS-CoV-2 positivity, 14% (95% CI 8–23%)–a subsequent history of positivity and 37% (95% CI 28–48%) were considered as false positive. Conclusion A substantial proportion of the inconclusive results should be considered as positive samples at the beginning or the end of the infection. However, the number of false-positive results is also significant and each patient’s result should be analyzed separately following the clinical symptoms and epidemiological data.