Specific detection of H5N1 avian influenza A virus in field specimens by a one-step RT-PCR assay

BMC Infectious Diseases - 2006
Lisa F. P. Ng1, Ian Barr2, Tung Nguyen3, Suriani Mohd Noor4, Rosemary Tan5, Lora V. Agathe1, Sanjay Gupta1, Hassuzana Khalil4, Thanh Long To3, Sharifah Syed Hassan4, Ee Chee Ren6
1Genome Institute of Singapore, 60 Biopolis Street, Genome, #02-01, 138672, Singapore
2WHO Influenza Centre, 45 Poplar Road, Parkville, Melbourne, 3052, Australia
3Virology Section, National Centre for Veterinary Diagnostics, Department of Animal Health, Vietnam
4Virology Section, Veterinary Research Institute, 31400, Ipoh, Perak, Malaysia
5Veredus Laboratories Pte Ltd, 83 Science Park Drive, #03-02A The Curie, Singapore Science Park 1, 118259, Singapore
6Department of Microbiology, Faculty of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2, 117597, Singapore

Tóm tắt

Abstract Background Continuous outbreaks of the highly pathogenic H5N1 avian influenza A in Asia has resulted in an urgent effort to improve current diagnostics to aid containment of the virus and lower the threat of a influenza pandemic. We report here the development of a PCR-based assay that is highly specific for the H5N1 avian influenza A virus. Methods A one-step reverse-transcription PCR assay was developed to detect the H5N1 avian influenza A virus. The specificity of the assay was shown by testing sub-types of influenza A virus and other viral and bacterial pathogens; and on field samples. Results Validation on 145 field specimens from Vietnam and Malaysia showed that the assay was specific without cross reactivity to a number of other infuenza strains as well as human respiratory related pathogens. Detection was 100% from allantoic fluid in H5N1 positive samples, suggesting it to be a reliable sampling source for accurate detection. Conclusion The assay developed from this study indicates that the primers are specific for the H5N1 influenza virus. As shown by the field tested results, this assay would be highly useful as a diagnostic tool to help identify and control influenza epidemics.

Từ khóa


Tài liệu tham khảo

Nicholson KG, Wood JM, Zambon M: Influenza. Lancet. 2005, 362: 1733-1745. 10.1016/S0140-6736(03)14854-4.

Zeitlin GA, Maslow MJ: Avian influenza. Curr Infect Dis Rep. 2005, 7: 193-199.

Cox NJ, Subbarao K: Influenza. Lancet. 1999, 354: 1277-1282. 10.1016/S0140-6736(99)01241-6.

Swayne DE, Suarez DL: Highly pathogenic avian influenza. Rev Sci Tech. 2000, 19: 463-482.

Chan PK: Outbreak of avian influenza A (H5N1) virus infection in Hong Kong in 1997. Clin Infect Dis. 2002, 34 (Suppl 2): S58-64. 10.1086/338820.

Peiris JSM, Yu WC, Leung CW, Cheung CY, Ng NF, Nicholls JM, Ng TK, Chan KH, Lai ST, Lim WL, Yuen YY, Guan Y: Re-emergence of fatal human influenza A subtype H5N1 disease. Lancet. 2004, 363: 617-619. 10.1016/S0140-6736(04)15595-5.

Yuen KY, Chan PK, Peiris JSM, Tsang DN, Que TL, Shortridge KF, Cheung PT, To WK, Ho ET, Sung R, Cheng AF: Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus. Lancet. 1998, 351: 467-471. 10.1016/S0140-6736(98)01182-9.

Centers for Disease Control and Prevention (CDC): Cases of influenza A (H5N1) – Thailand, 2004. Morb Mortal Wkly Rep. 2004, 53: 100-3.

Hien TT, Liem NT, Dung NT, San LT, Mai PP, Chau NV, Suu PT, Dong VC, Mai LTQ, Thi NT, Khoa DB, Phat LP, Troung NT, Long HT, Giang LT, Tho ND, Tien NTK, San LH, Tuan LV, Dolecek C, Thanh TT, Jong M, Schultsz C, Cheng P, Lim W, Horby P, Farrar J: Avian Influenza A (H5N1) in 10 patients in Vietnam. N Engl J Med. 2004, 350: 1179-1188. 10.1056/NEJMoa040419.

Lau L, Banks J, Aherne R, Brown IH, Dillon N, Collins RA, Chan K, Fung YW, Xing J, Yu ACH: Nucleic acid sequence-based amplification methods to detect avian influenza virus. Biochem Biophys Res Comm. 2003, 313: 336-342. 10.1016/j.bbrc.2003.11.131.

Lee M, Chang P, Shien J, Cheng M, Shieh HK: Identification and subtyping of avian influenza viruses by reverse transcription-PCR. J Virol Methods. 2001, 97: 13-32. 10.1016/S0166-0934(01)00301-9.

Payungporn S, Phakdeewirot P, Chutinimitkul S, Theamboonlers A, Keawcharoen J, Oraveerakul K, Amonsin A, Poovorawan Y: Single-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) for influenza A virus subtype H5N1 detection. Viral Immunol. 2004, 17: 588-593. 10.1089/vim.2004.17.588.

Thông tin
Thông tin xuất bản

BMC Infectious Diseases

Thông tin tác giả