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Loop-Mediated Isothermal Amplification Assay for Peste des Petits Ruminants Virus Detection and its Comparison with RT-PCR
 
Yasmin Niamat1, Mudasser Habib1*, Mudassar Mohiuddin2 and Muhammad Salahuddin Shah1
 
1Animal Science Division, NIAB, Faisalabad; 2Guangdong Haid Institute of Animal Husbandry and Veterinary, Guangzhou, Guangdong, 511400, China
*Corresponding author: mudasserhabib@yahoo.com
 

Abstract   

Peste des petits ruminant (PPR) represents an acute, highly contagious, transboundary viral disease of goats (predominantly) and sheep. The disease is endemic in Pakistan with annual losses of 20.5 billion rupees. The objective of the study was to optimize the rapid, efficient and more sensitive diagnostic tool for disease outbreak investigation and field surveillance. Accordingly, single tube loop mediated isothermal amplification (LAMP) technique was performed by incubating virus and/or suspected clinical samples at 58°C for 60min, and it was compared with conventional reverse transcription polymerase chain reaction (RT-PCR). Based on sequence analysis, 2 primer pairs were used, named as NP3/NP4 and NPx/NPy (this study) for RT-PCR to target the nucleoprotein gene and three sets of primers namely, F3/B3, FIP/BIP, F loop/B loop for RT-LAMP to target the matrix gene. The NP3/NP4 primers performed better compared with our designed NPx/NPy primers in RT-PCR. Twenty-five clinical samples were evaluated by LAMP assay, and the results were found consistent with those of conventional RT-PCR. LAMP primers showed high specificity and had no cross reaction with other animal viruses i.e. FMDV and IBDV. The sensitivity of LAMP assay was 10-fold higher than conventional RT-PCR. As a simple, inexpensive and accurate detection method, RT-LAMP assay is well suited for rapid clinical diagnosis of PPR. Therefore, it can easily replace RT-PCR for disease investigation and surveillance of PPRV in field conditions especially in developing countries like Pakistan.

To Cite This Article: Niamat Y, Habib M, Mohiuddin M and Shah M, 2018. Loop-Mediated Isothermal Amplification Assay for Peste des Petits Ruminants Virus Detection and its Comparison with RT-PCR. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2018.106

 
   

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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