Abstract
A specific and sensitive two-step TaqMan real-time PCR has been developed for rapid diagnosis of caprine arthritis-encephalitis virus (CAEV) infection by using a set of specific primers and a TaqMan probe targeting a highly conserved region within the gene encoding the viral capsid protein (CA). The assay successfully detected CAEV proviral DNA in total DNA extracts originating from cell culture, whole blood samples and isolated PBMCs, with a lower detection limit of 102 copies and a linear dynamic range of 105 to 1010 copies/ml. There was no cross-reaction with other animal viruses (e.g., goat pox virus, bovine leukemia virus, bovine mucosal disease virus, swine influenza virus and Nipah virus). When applied in parallel with serological AGID and conventional PCR for detection of CAEV in field samples, this assay exhibited a higher sensitivity than these traditional methods, and 7.8 % of the 308 specimens collected in the Shanxi and Tianjin regions of China from 1993 to 2011 were found to be positive. Thus, the TaqMan qPCR assay provides a fast, specific and sensitive means for detecting CAEV proviral DNA in goat specimens and should be useful for large-scale detection in eradication programs and epidemiological studies.
Highlights
Caprine arthritis-encephalitis virus (CAEV) is a member of the genus Lentivirus, family Retroviridae [4] and induces persistent and progressive degenerative inflammatory disease in infected goats [20]
To determine the sensitivity of the TaqMan qPCR assay, the recombinant plasmid pGEM-T-capsid protein (CA) was prepared in tenfold serial dilutions, and 1 lL of each serially diluted recombinant plasmid sample was used as template
caprine arthritis-encephalitis virus (CAEV) control has remained a big challenge for the goat industry, as prophylactic vaccinations do not induce antibodies that result in efficient viral clearance and provide protection against arthritis [11]
Summary
Caprine arthritis-encephalitis virus (CAEV) is a member of the genus Lentivirus, family Retroviridae [4] and induces persistent and progressive degenerative inflammatory disease in infected goats [20]. CAEV infection is one of the most destructive and economically important viral diseases of the goat industry and is spread throughout many countries of the world, including the United States (31 %) [30], Norway (86 %) [22], Great Britain (54.5 %) [32], Switzerland (26.9 %) [3], Spain (20.6 %) [31], Poland (12.1 %) [15], Italy (6.58 %) [11], Japan (63.3 %) [14], Mexico (56.8 %) [33], Brazil (35 %) [18], Jordan (23.2 %) [1], Korea (2.73 %) [23] and China (0.2 %–30 %) [26]. Economic losses attributed to CAEV infection are considerably adverse in countries with intensive animal husbandry, with 5 %–10 % goats reported to be culled annually due to arthritis, and the decrease in milk production in infected does was estimated to be 10 %–15 % in Switzerland [24]. In udder halves with intramammary infection, milk SCC (somatic cell count) was significantly increased [17]
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