Rapid detection of fowl adenovirus field samples using loop-mediated isothermal amplification assay

M PARTHIBAN; K S SHALINI; KURUNCHI C DIVYA; K KUMANAN; K S AARTHI

doi:10.56093/ijans.v84i1.37295

Authors

M PARTHIBAN Madras Veterinary College, Chennai, Tamil Nadu 600 007 India
K S SHALINI Madras Veterinary College, Chennai, Tamil Nadu 600 007 India
KURUNCHI C DIVYA Madras Veterinary College, Chennai, Tamil Nadu 600 007 India
K KUMANAN Madras Veterinary College, Chennai, Tamil Nadu 600 007 India
K S AARTHI Madras Veterinary College, Chennai, Tamil Nadu 600 007 India

https://doi.org/10.56093/ijans.v84i1.37295

Keywords:

Fowl adenovirus, Hexon gene, Loop mediated isothermal amplification assay, Polymerase chain reaction, Real time PCR

Abstract

Loop-mediated isothermal amplification was employed for the detection of fowl adenoviruses as a simple, rapid and field based diagnostic assay. Out of 134 field samples screened, 28 samples were positive by both PCR and LAMP assay. The LAMP assay primers were found to be highly specific and it detected only fowl adenovirus samples and it did not react with other avian viruses like Marek’s disease virus and avian leucosis virus. Conventional PCR detected 100ug level of DNA whereas LAMP assay detected up to 10ng level of DNA. It clearly indicated that LAMP assay was 100-times more sensitive than conventional PCR. The specificity of LAMP assay was confirmed by sequencing of the LAMP amplified products and real time PCR. The LAMP positive reaction can be easily visualized under UV trans- illuminator by the addition of SYBR green dye. Thus, LAM Passay proves to be a sensitive, rapid, less-laborious and cost- effective technique for the diagnosis of fowl adenoviruses in field conditions.

Downloads

Download data is not yet available.

References

Ashok K R, Praveen K B, Rajeev K R, Annie M and Inderjeet K. 2010. Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid and Inexpensive Detection of Cytomegalovirus DNA in Vitreous Specimens from Suspected Cases of Viral Retinitis. Journal of Clinical Microbiology 48: 2050–52.

Das S, Pingle M R, Mun˜oz-Jorda´n J, Rundell M S, Rondini S, Granger K, Chang G J, Kelly E, Spier E G, Larone D, Spitzer E, Barany F and Golightly L M. 2008. Detection and Serotyping of Dengue Virus in Serum Samples by Multiplex Reverse Transcriptase PCR-Ligase Detection Reaction Assay. Journal of Clinical Microbiology 46: 3276–84.

Ganesh K, Suryanarayana V V and Raghavan R. 2002. Detection of fowl adenovirus associated with hydropericardium hepatitis syndrome by a polymerase chain reaction. Veterinary Research Communications 26: 73–80.

Soliman H and El-Matbouli M. 2010. Loop mediated isothermal amplification combinedwith nucleic acidnlateral flow strip for diagnosis of cyprinid herpes virus-3. Molecular and Cellular Probes 24: 38–43.

Norrby E. 1969. The relationship between the soluble antigens and the virion of adenovirus type3. IV. Immunological complexity of soluble components. Journal of Virolgy 37: 565–76.

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N and Hase T. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acid Research 28: E 63.

Parthiban M, Divya K C, Kumanan K and Bargavi D S. 2012. A rapid and highly reliable field based diagnostic assay for detection of Canine Parvovirus. Acta Virologica 56: 71– 74.

Parida M, Posadas G, Inoue S, Hasebe F and Morita K.2004. Real- time reverse transcription loop-mediated isothermal amplification for rapid detection of West Nile virus. Journal of Clinical Microbiology 42: 257–63.

Steer P A, Kirkpatrick N C, O’Rourke D and Noormohammadi A H. 2009. Classification of Fowl Adenovirus Serotypes by Use of High-Resolution Melting-Curve Analysis of the Hexon Gene Region. Journal of Clinical Microbiology 47: 311–21.

Raue R and Hess M. 1998. Hexon based PCRs combined with restriction enzyme analysis for rapid detection and differentiation of fowl adenoviruses and egg drop syndrome virus. Journal of Virological Methods 73: 211–17.

Saito R, Misawa Y, Moriya K, Koike K, Ubukata K and Okamura N. 2005. Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae. Journal of Medical Microbiology 54: 1037–41.

Shanthi J, Chung Y C, Barr I, Kwok H C, Honglin C, Yi G, Malik P J S and Leo Poon L M. 2007. Loop-mediated isothermal amplification for Influenza A (H5N1) Virus. Emerging Infectious Diseases 13: 899–901.

Winterfield R W, Fadly, A M and Gallina A M. 1973. Adenovirus infection and disease. 1. some characteristics of an isolate from chickens in Indiana. Avian Diseases 17: 334–42.

Xue C, Zhang Y, Zhou Q, Xu C, Li X and Cao Y. 2009. Rapid detection of Infectious bursal disease virus by reverse transcription loop-mediated isothermal amplification assay. Journal of Veterinary Diagnostic Investigations 21: 841– 43.

Pandey B D, Poudel A, Yoda T, Tamaru A, Oda N, Fukushima Y, Lekhak B, Risal B Acharya B, Sapkota B, Nakajima C, Taniguchi T, Phetsuksiri B and Suzuki Y. 2008. Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients. Journal of Medical Microbiology 57: 439–43.

Yi P, Zhixun X, Jiabo L, Yaoshan P, Xianwen D, Zhiqin X, Liji X, Qing F, Jiaxun F and Mazhar I K. 2011. Visual detection of H3 sub type avian influenza viruses by reverse transcription loop-mediated isothermal amplification assay. Journal of Virology 8: 337–47.

Xie Z, Tang Y, Fan Q, Liu J, Pang Y, Deng X, Xie Z, Peng Y, Xie L and Khan M I. 2011. Rapid detection of Group I Avian Adenoviruses by a Loop-mediated Isothermal Amplification. Avian Diseases 55: 575–79.