tetano
Editor, Senior Moderator
J Clin Microbiol. 2013 Oct 23. [Epub ahead of print]
Newly emerging mutations in the matrix genes of the human influenza A(H1N1)pdm09 and A(H3N2) viruses reduce the detection sensitivity of the real-time RT-PCR assay.
Yang JR, Kuo CY, Huang HY, Wu FT, Huang YL, Cheng CY, Su YT, Chang FY, Wu HS, Liu MT.
Source
Centers for Disease Control, Taipei, Taiwan, ROC.
Abstract
New variants of the influenza A(H1N1)pdm09 and A(H3N2) viruses were detected in Taiwan between 2012 and 2013. Some of these variants were not detected in clinical specimens using a common real-time reverse transcription polymerase chain reaction (RT-PCR) assay that targeted the conserved regions of the viral Matrix (M) genes. Analysis of the M gene sequences of the new variants revealed that several newly emerging mutations were located in the regions where the primers/probes of the real-time RT-PCR assay bind; these included three mutations (G225A, T228C, and G238A) in the H1N1pdm09 virus, as well as one mutation (C163T) in the H3N2 virus. These accumulated mismatch mutations, together with the previously identified C154T mutant of the H1N1pdm09 virus and the C153T and G189T mutants of the H3N2 virus, result in a reduced detection sensitivity of the real-time RT-PCR assay. To overcome the loss of sensitivity due to mismatch mutations, we established a real-time RT-PCR assay using degenerate nucleotide bases in both the primers and probe and successfully increased the sensitivity of the assay to detect the circulating variants of the human influenza A viruses. Our observations highlight the importance of the simultaneous use of different gene-targeting real-time RT-PCR assays in the clinical diagnosis of influenza.
PMID:
24153120
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24153120
Newly emerging mutations in the matrix genes of the human influenza A(H1N1)pdm09 and A(H3N2) viruses reduce the detection sensitivity of the real-time RT-PCR assay.
Yang JR, Kuo CY, Huang HY, Wu FT, Huang YL, Cheng CY, Su YT, Chang FY, Wu HS, Liu MT.
Source
Centers for Disease Control, Taipei, Taiwan, ROC.
Abstract
New variants of the influenza A(H1N1)pdm09 and A(H3N2) viruses were detected in Taiwan between 2012 and 2013. Some of these variants were not detected in clinical specimens using a common real-time reverse transcription polymerase chain reaction (RT-PCR) assay that targeted the conserved regions of the viral Matrix (M) genes. Analysis of the M gene sequences of the new variants revealed that several newly emerging mutations were located in the regions where the primers/probes of the real-time RT-PCR assay bind; these included three mutations (G225A, T228C, and G238A) in the H1N1pdm09 virus, as well as one mutation (C163T) in the H3N2 virus. These accumulated mismatch mutations, together with the previously identified C154T mutant of the H1N1pdm09 virus and the C153T and G189T mutants of the H3N2 virus, result in a reduced detection sensitivity of the real-time RT-PCR assay. To overcome the loss of sensitivity due to mismatch mutations, we established a real-time RT-PCR assay using degenerate nucleotide bases in both the primers and probe and successfully increased the sensitivity of the assay to detect the circulating variants of the human influenza A viruses. Our observations highlight the importance of the simultaneous use of different gene-targeting real-time RT-PCR assays in the clinical diagnosis of influenza.
PMID:
24153120
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24153120