Evaluation of filter paper to transport oro/nasopharyngeal samples to detect SARS-CoV-2 by RT-qPCR.

PURPOSE: To evaluate filter paper as a means to transport oro/nasopharyngeal samples from laboratories with few resources for SARS-CoV-2 detection by RT-qPCR in a central laboratory that usually performs this technique as routine. METHODS: A total of 40 specimens were evaluated in parallel by RT-qPCR carried out after RNA extraction using two different protocols: direct RNA extraction (Protocol A - reference method) and RNA extraction after impregnation in filter paper (Protocol B). RESULTS: The RT-qPCR for SARS-CoV-2 using Protocol B presented 97.22% (35/36) of agreement for SARS-CoV-2-positive samples when compared to the reference method (Protocol A), even for specimens with low viral load (increased Ct values). Noteworthy, three clinical specimens which were categorized as inconclusive by Protocol A presented amplification of both N1 and N2 targets using Protocol B, presenting positive results for SARS-CoV-2. CONCLUSION: The use of filter paper to transport oro/nasopharyngeal clinical samples presented very satisfactory results to detect SARS-CoV-2 by RT-qPCR. In addition, it proved to be a feasible and sensitive approach, being able to generate the detection of SARS-CoV-2 even at low concentrations, without presenting false-negative results.

Novel mutations in the resistome of a new sequence type (ST262) of clarithromycin resistant Mycobacterium abscessus subsp. massiliense.

OBJECTIVES: The aim of this study was to evaluate the relationship between antimicrobial susceptibility and genotype of a Mycobacterium abscessus subsp. massiliense isolate obtained from the respiratory tract of a patient in southern Brazil. METHODS: The isolate (named Myco1POA) was submitted to whole-genome sequencing using an Illumina MiSeq platform. Data were analysed using Trim Galore!, SPAdes Genome Assembler, Geneious and BioEdit software. Multilocus sequence typing (MLST) was performed by in silico analysis of seven housekeeping genes according to the Institut Pasteur database. The antimicrobial susceptibility profile was determined by broth microdilution according to Clinical and Laboratory Standards Institute (CLSI) guidelines. RESULTS: Several mutations in genes related to antimicrobial resistance were identified in Myco1POA. MLST indicated that the isolate belonged to a novel sequence type (ST), which was designated ST262. Phenotypic susceptibility and genotypic findings were concordant, except for clarithromycin [erm(41) and rrl genes]. CONCLUSION: Here we describe the genome sequence of M. abscessus subsp. massiliense Myco1POA identified as a novel sequence type (ST262) and indicate possible new gene mutations leading to clarithromycin resistance.