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. 2002 Apr 10;16(3):163–166. doi: 10.1002/jcla.10036

Evaluation of a single‐step multiplex RT‐PCR for influenza virus type and subtype detection in respiratory samples

Saibal K Poddar 1,✉, Rowena Espina 1, David P Schnurr 2
PMCID: PMC6807991  PMID: 11968055

Abstract

A single‐step multiplex reverse transcription‐polymerase chain reaction (RT‐PCR) for the simultaneous detection of influenza virus type and subtypes was evaluated for its use in clinical specimens. One part of each specimen was tested using an established standard culture and/or monoclonal antibody‐based immunofluorescence assay method, and the other part was tested by the multiplex RT‐PCR method for the presence or absence of the influenza virus, and its type and subtype. Sixty‐seven specimens were examined. The results revealed a strong agreement between the data obtained by the established method and those obtained by the multiplex RT‐PCR. J. Clin. Lab. Anal. 16:163–166, 2002. © 2002 Wiley‐Liss, Inc.

Keywords: influenza virus, multiplex PCR, typing, subtyping

REFERENCES

  • 1. Webster RG. 1998. Influenza: an emerging disease. Emerg Infect Dis 4:436–441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Bao‐Lan L, Webster RG, Brown LE. 1983. Nerome K: heterogeneity of influenza B viruses. Bull World Health Organ 61:681–687. [PMC free article] [PubMed] [Google Scholar]
  • 3. Brammer L, Fukuda K, Arden N, et al. 1997. Influenza surveillance, United States, 1992–93 and 1993–94. Morbid Mortal Wkly Rep CDC Surveill Summ 46:1–12. [PubMed] [Google Scholar]
  • 4. Merchant GL, Mower WR, Talan DA. 1997. Update on emerging infections from the Centers for Disease Control and Prevention. Influenza: ED considerations for the 1997–98 season. Ann Emerg Med 30:692–694. [DOI] [PubMed] [Google Scholar]
  • 5. Class EC, Osterhaus AD, van‐Beek R, et al. 1998. Human influenza A H5N1 virus related to a highly pathogenic avian virus. Lancet 351:472–477. [DOI] [PubMed] [Google Scholar]
  • 6. Harmon MW. 1992. Influenza viruses In: Lennette EH, editor. Laboratory diagnosis of viral infections. 2nd ed. New York: Marcel Dekker; p 515–534. [Google Scholar]
  • 7. Herrmann B, Larsson C, Zweygberg BW. 2001. Simultaneous detection and typing of influenza viruses A and B by a nested reverse transcription‐PCR: comparison to virus isolation and antigen detection by immunofluorescence and optical immunoassay (FLU OIA). J Clin Microbiol 3:134–138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. Poddar SK, Sawyer MH, Connor JD. 1997. Optimized PCR amplification of influenza A virus RNA using Tth DNA polymerase incorporating uracil N glycosylase (UNG) in a single tube reaction. J Clin Lab Anal 11:323–327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Poddar SK, Sawyer MH, Connor JD. 1998. Evaluation of PCR assays in presence of antibody to thermostable DNA polymerases for detection of microbial agents: avoiding false negative results for specimens containing low titer agent. J Clin Lab Anal 12:238–241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. Cherian T, Bobo L, Steinhoff MC, Karron RA, Yolken RH. 1994. Use of PCR‐enzyme immunoassay for identification of influenza A virus matrix RNA in clinical samples negative for cultivable virus. J Clin Microbiol 32:623–628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. Atmar RL, Baxter BD, Dominguez EA, Taber LH. 1996. Comparison of reverse transcription‐PCR with tissue culture and other rapid diagnostic assays for detection of type A influenza virus. J Clin Microbiol 34:2604–2606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Wright KE, Wilson GAR, Novosad D, Dimock C, Tan D, Weber JM. 1995. Typing and subtyping of influenza viruses in clinical samples by PCR. J Clin Microbiol 33:1180–1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. Shimizu H, Watanabe S, Imai M. 1997. Rapid detection of influenza A (AH1, AH3) and B by nested‐polymerase chain reaction. J Japn Assoc Infect Dis 71:522–526. [DOI] [PubMed] [Google Scholar]
  • 14. Poddar SK. 2002. Influenza virus types and subtypes detection by single step single tube multiplex reverse transcription‐polymerase chain reaction (RT‐PCR) and agarose gel electrophoresis. J Virol Methods 99:63–70. [DOI] [PubMed] [Google Scholar]
  • 15. Waecker NJ Jr, Shope TR, Weber PA, Buck ML, Domingo RC, Hooper DG. 1993. The rhino‐probe nasal curette for detecting respiratory syncytial virus in children. Pediatr Infect Dis J 4:326–329. [DOI] [PubMed] [Google Scholar]
  • 16. Schmidt NJ, Ota M, Gallo D, Fox VL. 1982. Monoclonal antibodies for rapid strains specific identification of influenza virus isolates. J Clin Microbiol 16:763–765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Schnurr D, Devlin V, Klimov A, Povinelli L, Hendry RM. 1998. H1 and H3 specific monoclonal antibody for subtyping influenza virus (abstr). M23. In: Proceedings of the 14th Annual Virology Symposium.
  • 18. Poddar SK, Sawyer MH, Connor JD. 1998. Effect of inhibitors in clinical specimens on Taq and Tth DNA polymerase‐based PCR amplification of influenza A virus. J Med Microbiol 47:1–5. [DOI] [PubMed] [Google Scholar]

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