Kombinasi One-step Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) dan Nucleic Acid Lateral Flow (NALF) sebagai Metode Deteksi Gen env-tm Virus Jembrana Strain Tabanan 1987

https://doi.org/10.22146/jsv.41146

Asmarani Kusumawati(1*), Fatimah Fatimah(2)

(1) Fakultas Kedokteran Hewan Universitas Gadjah Mada, Yogyakarta
(2) Sekolah Pascasarjana Universitas Gadjah Mada, Yogyakarta
(*) Corresponding Author

Abstract


Jembrana disease is an acute viral disease  in Bali cattle that have short incubation period, high mortality and morbidity rate. Early diagnostic methods are needed  to prevent the further spreading of this disease. In this study, we combined One-step RT-PCR  and  NALF  methods to detect env-tm gene of Jembrana virus Tabanan 1987 strain. Viral RNA isolated from a spleen of the infected cattle was used as the template. One-step RT-PCR procedure was performed using oligoprobes  labeled digoxigenin and  right primer sequence that were designed using the primer3plus program based  on the conserves region of env-tm gene from NCBI database. The product of one-step RT-PCR was tested using NALF  method  instead of  electrophoresis. Positive result was shown by the appearance of dark lines on the test line of digoxigenin in NALF dipstick device.


Keywords


RT-PCR; NALF; Jembrana disease virus Tabanan 1987 strain; env-tm gene; digoxigenin

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References

Andrade, T. P. D, and Lightner, D. V. 2009. Development of a method for detection of infectious   myonecrosis virus by reverse-transcription loop- mediated isothermal amplification and    nucleic acid lateral flow hybrid assay. Journal of Fish Disease 32.

 

Chadwick, B. J., Coelen, R. J., Wilcox, G E., Sammels, L. M., Kertayadnya, G. 1995.

Nucleotide sequence analysis of Jembrana disease virus: a bovine lentivirus associated with an acute disease syndrome. Journal of  General  Virology 76: 1637 – 1650.

 

Dayanghirang, J. A. and Masanori, M. 1993. Quantitative determination of PCR products by colorimetric method. Jpn. J. Leprosy 62: 21-27.

 

Desport, M. and Joshua, L. 2010. Jembrana Disease Virus: host response, viral dynamics, and disease control. Current HIV  Research 8: 53-65.

 

Ditcham, W. G. F., Joshua, R. L., Robert, J. D., Nining, H., Graham, E. W., Moira, D. 2009. Vaccination reduces the viral load and the risk of transmission of Jembrana disease virus in Bali cattle. Virology 386: 317-324.

 

Dos Santos, C. F., Vivien, T. S., Maria, A. D. A. M., Daniela., N. S., Andrew, S. G. 2004. Reverse transcription and polymerase chain reaction: principles and aplications in dentistry. Journal  Applied Oral Science 12:1-11.

 

Hartaningsih, N., Wilcox, G.E., Dharma, D.M., Soetrisno, M., 1993. Distribution of Jembrana disease in cattle in Indonesia. Veterinary  Microbiology 38: 23–29.

 

Kamphee, H., Angkana, C., Therdsak, P., Natpapas, W., Airin, K., Tararaj, D. 2015. Rapid Molecular Detection of Multidrug-Resistant Tuberculosis by PCR-Nucleic Acid Lateral Flow Immunoassay. PloS ONE 10: e0137791. doi: 10.1371/journal.pone.0137791.

 

Kertayadnya, G., Wilcox, G. E., Soeharsono, S., Hartaningsih, N., Coelen, R. J., Cook, R. D., Collins, M. E., Brownlie, J. 1993. Characteristics of a retrovirus associated with Jembrana disease in Bali cattle. Journal of  General  Virology 74: 1765 – 1778.

 

Kusumawati, A., Pratiwi, R., Astuti, P., Hamid, P. H. 2010. Characterization of envelope-transmembrane gene of jembrana disease virus tabanan 1995 isolate. Indonesian Journal of Biotechnol 15: 15 – 19.

 

Kusumawati, A., Tenri, A. W., Rizqa, F. P., Basofi, A. M., Isabellina, D. T. 2014. The structure and function of jembrana disease virus genom. Journal Infection and Molecular Biology 2: 26-29.

 

Rohrman, B. A., Leautaud, V., Molyneux, E., Richards-Kortum, R. R. 2012. A Lateral Flow Assay for Quantitative Detection of Amplified HIV-1 RNA. PLoSONE 7: e45611. doi:10.1371/journal.pone.0045611.

 

Haridas, T. K., Thiruvengadam, S., Bishor, V. I. 2014. Development of a PCR Based Nucleic Acid Lateral Flow Assay Device for Detection of Mycobacterium Tuberculosis Complex. International Journal of PharmTech Research 6: 1695-1702.

 

Posthuma-Trumpie, GA. Korf, J. And van Amerongen, A.2008.  Lateral flow (immuno)assay:its strengths, weaknesses, opportunities and threats. A literature survey. Analytical and Bioanalytical Chemistry 393: 569–582.

 

Ramachandran, S. 1996. Early observations and research on jembrana disease in Bali and other Indonesian Islands. ACIAR Proceeding 75: 5-9.

 

Shiao, Y. H. 2003. A new reverse transcription-polymerase chain reaction method for

accurate quantification. BMC Biotechnology 3: 22-34.

 

Soeharsono, S., Hartaningsih, N., Soetrisno, M., Kertayadnya, G., Wilcox, G.E. 1990. Studies of experimental Jembrana disease in Bali cattle. I. Transmission and persistence of the infectious agent in ruminants and pigs, and resistance of recovered cattle to re-infection. Journal of Comparative Pathology 103: 49-59.

 

Soeharsono, S., Wilcox, G. E., Putra, A. A, Hartaningsih, N., Sulistyana, K., Tenaya, M. 1995. The transmission of Jembrana disease, a lentivirus disease of  Bos javanicus cattle. Epidemiology and Infection 115: 367-374.

 

Wacker, M. J., and Michael, P. G. 2005. Analysis of One-Step and Two-Step Real-Time RT-PCR Using SuperScript III. Journal of  Biomolecular Technology 16:266–271.

 

Wilcox, G., 1997. Jembrana disease. Australian Veterinary Journal 7: 492–493.



DOI: https://doi.org/10.22146/jsv.41146

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