Giuseppe
Emeritus
J Clin Microbiol. 2008 Dec 10. [Epub ahead of print]
A Multiplex Assay for Simultaneously Typing and Subtyping Influenza Viruses on an Electronic Microarray.
Huang Y, Tang H, Duffy S, Hong Y, Norman S, Ghosh M, He J, Bose M, Henrickson KJ, Fan J, Kraft AJ, Weisburg WG, Mather EL. - Nanogen, Inc., San Diego, California 92121; and Department of Pediatrics, Midwest Respiratory Virus Program, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
We report on the use of an electronic microarray to simultaneously type influenza A and B and distinguish influenza A subtypes H1N1 and H3N2 from the potentially pandemic avian subtype H5N1.
The assay targets seven genes, including H1, H3, H5, N1, N2 genes of influenza A virus as well as the matrix protein M1 gene of influenza A and the nonstructural protein NS gene of influenza B.
By combining a two-step reverse transcription multiplex PCR with typing and subtyping on the electronic microarray, the assay achieved analytical sensitivity of 10(2)-10(3) copies of transcripts for each of the genes.
The assay correctly typed and subtyped fifteen different influenza viral isolates, including two influenza B, five A/H1N1, six A/H3N2, as well as two A/H5N1 isolates.
In addition, the assay correctly identified 8 out of 10 diluted, archived avian specimens with complete typing and subtyping information and 2 specimens with partial subtyping information.
In a study of one hundred and forty-six human clinical specimens that had been previously shown to be positive for influenza or another respiratory virus, the assay showed a clinical sensitivity of 96 % and clinical specificity of 100%.
The assay is a rapid, accurate, user-friendly method for simultaneously typing and subtyping influenza viruses.
PMID: 19073867 [PubMed - as supplied by publisher]
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A Multiplex Assay for Simultaneously Typing and Subtyping Influenza Viruses on an Electronic Microarray.
Huang Y, Tang H, Duffy S, Hong Y, Norman S, Ghosh M, He J, Bose M, Henrickson KJ, Fan J, Kraft AJ, Weisburg WG, Mather EL. - Nanogen, Inc., San Diego, California 92121; and Department of Pediatrics, Midwest Respiratory Virus Program, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
We report on the use of an electronic microarray to simultaneously type influenza A and B and distinguish influenza A subtypes H1N1 and H3N2 from the potentially pandemic avian subtype H5N1.
The assay targets seven genes, including H1, H3, H5, N1, N2 genes of influenza A virus as well as the matrix protein M1 gene of influenza A and the nonstructural protein NS gene of influenza B.
By combining a two-step reverse transcription multiplex PCR with typing and subtyping on the electronic microarray, the assay achieved analytical sensitivity of 10(2)-10(3) copies of transcripts for each of the genes.
The assay correctly typed and subtyped fifteen different influenza viral isolates, including two influenza B, five A/H1N1, six A/H3N2, as well as two A/H5N1 isolates.
In addition, the assay correctly identified 8 out of 10 diluted, archived avian specimens with complete typing and subtyping information and 2 specimens with partial subtyping information.
In a study of one hundred and forty-six human clinical specimens that had been previously shown to be positive for influenza or another respiratory virus, the assay showed a clinical sensitivity of 96 % and clinical specificity of 100%.
The assay is a rapid, accurate, user-friendly method for simultaneously typing and subtyping influenza viruses.
PMID: 19073867 [PubMed - as supplied by publisher]
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