利用逆转录聚合酶链反应和互补脱氧核糖核酸：核糖核酸原位杂交技术对蛇副黏病毒感染进行分子诊断

Molecular diagnosis of paramyxovirus infection in snakes using reverse transcriptase-polymerase chain reaction and complementary deoxyribonucleic acid:ribonucleic acid in situ hybridization.

作者信息

Sand Matthew A, Latimer Kenneth S, Gregory Christopher R, Rakich Pauline M, Jacobson Elliott, Pennick Kate E

机构信息

Honors Microbiology Program, University of Georgia, Athens, GA 30602, USA.

出版信息

J Vet Diagn Invest. 2004 Sep;16(5):442-8. doi: 10.1177/104063870401600514.

DOI:10.1177/104063870401600514

PMID:15460330

Abstract

Identification of ophidian paramyxovirus (OPMV) nucleic acid was accomplished in 11 of 14 snakes by a reverse transcriptase-polymerase chain reaction (RT-PCR) assay that detected a 153-bp region of the OPMV genome in total RNA extracted from paraffin-embedded tissues and cell culture. The RT-PCR protocol amplified a portion of the OPMV RNA genome, producing a 153-bp complementary DNA (cDNA) product from both fresh and paraffin-embedded tissue samples. In addition, cDNA:RNA in situ hybridization localized OPMV in formalin-fixed, paraffin-embedded tissue specimens to specific tissues and cells. This latter technique increased the degree of specificity with which a diagnosis of OPMV could be made.

摘要

通过逆转录聚合酶链反应（RT-PCR）检测法，在14条蛇中的11条身上鉴定出了蛇副粘病毒（OPMV）核酸。该检测法可从石蜡包埋组织和细胞培养物中提取的总RNA中检测出OPMV基因组的一个153碱基对区域。RT-PCR方案扩增了OPMV RNA基因组的一部分，从新鲜组织样本和石蜡包埋组织样本中均产生了一个153碱基对的互补DNA（cDNA）产物。此外，cDNA:RNA原位杂交将OPMV定位在福尔马林固定、石蜡包埋的组织标本中的特定组织和细胞中。后一种技术提高了诊断OPMV的特异性程度。