[A specific PCR assay and a nested PCR assay for the screening of ovine pulmonary adenomatosis].
Ovine pulmonary adenomatosis (OPA) is a naturally occurring contagious lung tumor of sheep which was caused by an exogenous retrovirus of sheep, jaagsiekte retrovirus (JSRV). Although no specific circulating antibodies against the virus coud be detected in infected sheep, exogenous JSRV proviral DNA sequences (exJSRV) and JSRV RNA transcripts could be detected in lung tumors, lymphoreticular system and peripheral blood mononuclear cells (PBMC) from sheep affected by OPA. The sheep genome carried 15 to 20 copies of endogenous retrovirus loci (enJSRV) that were similar to JSRV in structural genes but the divergene in U3. Therefore, primers specific for the U3 sequences of exJSRV were designed for the specific PCR and nested PCR (n-PCR). Sensitivity between specific PCR assay and n-PCR assay was compared by using serial dilutions of positive plasmid pJSRV-LTR in a background of 700ng sheep genome DNA. Sensitivity of n-PCR was ten-fold higher than specific PCR. The n-PCR was only available in blood test for detection of JSRV infected sheep and might be useful in epidemiological studies and disease control of OPA.
Luo
Jun-Rong
JR
College of Animal Science and Animal Medicine, Inner Mongolia Agricultural University, Huhhot. junrluo@126.com
Liu
Xiao-Hui
XH
Yu
Qun-Ying
QY
Zhang
Shu-Qing
SQ
Zhou
Jian-Hua
JH
Ma
Xue-En
XE
D000818:Animals; D001483:Base Sequence; D019411:Clinical Laboratory Techniques; D004279:DNA, Viral / Q000032:analysis*; D020077:Endogenous Retroviruses / Q000235:genetics; D022141:Jaagsiekte sheep retrovirus / Q000235:genetics*; D008175:Lung Neoplasms / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011648:Pulmonary Adenomatosis, Ovine / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012756:Sheep; D012757:Sheep Diseases / Q000821:virology; D014776:Virus Cultivation
D004279:DNA, Viral