Caprine arthritis encephalitis virus detection in blood by loop-mediated isothermal amplification (LAMP) assay targeting the proviral gag region

Michelle M Balbin; Lawrence P Belotindos; Nancy S Abes; Claro N Mingala

doi:10.1016/j.diagmicrobio.2013.12.012

Caprine arthritis encephalitis virus detection in blood by loop-mediated isothermal amplification (LAMP) assay targeting the proviral gag region

Diagn Microbiol Infect Dis. 2014 May;79(1):37-42. doi: 10.1016/j.diagmicrobio.2013.12.012. Epub 2014 Jan 7.

Authors

Michelle M Balbin¹, Lawrence P Belotindos¹, Nancy S Abes¹, Claro N Mingala²

Affiliations

¹ Animal Health Unit, Philippine Carabao Center National Headquarters and Gene Pool, Science City of Munoz 3120, Nueva Ecija, Philippines.
² Animal Health Unit, Philippine Carabao Center National Headquarters and Gene Pool, Science City of Munoz 3120, Nueva Ecija, Philippines. Electronic address: cnmingala@hotmail.com.

PMID: 24630755
DOI: 10.1016/j.diagmicrobio.2013.12.012

Abstract

Caprine arthritis encephalitis virus (CAEV), of the genus Lentivirus of the Retroviridae family, causes persistent disease, which is characterized by polyarthritis and mastitis in adult goats and progressive paresis (leukoencephalomyelitis) in kids. A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of CAEV in blood samples. Species-specific primers amplifying the gag gene region in the provirus were used for the detection of CAEV. The LAMP assay result was obtained 30 min after incubation on a constant temperature at 63 °C in a heat block. Resulting amplicons were visualized by addition of SYBR green dye after the reaction and checked by agarose gel electrophoresis. The sensitivity of LAMP assay was evaluated by comparing the result with the nested polymerase chain reaction. Based on the experiments, the result of the assay indicated a rapid and sensitive test for the detection of CAEV.

Keywords: Caprine arthritis encephalitis virus; Gag gene region; Loop-mediated isothermal amplification; Nested-PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Arthritis-Encephalitis Virus, Caprine / genetics*
Arthritis-Encephalitis Virus, Caprine / isolation & purification*
Electrophoresis, Agar Gel
Gene Products, gag / genetics*
Goats
Lentivirus Infections / diagnosis
Lentivirus Infections / virology*
Nucleic Acid Amplification Techniques / methods*

Substances

Gene Products, gag