Molecular diagnosis of paramyxovirus infection in snakes using reverse transcriptase-polymerase chain reaction and complementary deoxyribonucleic acid:ribonucleic acid in situ hybridization.
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.
Honors Microbiology Program, University of Georgia, Athens, GA 30602, USA.
Matthew A
MA
Sand
Kenneth S
KS
Latimer
Christopher R
CR
Gregory
Pauline M
PM
Rakich
Elliott
E
Jacobson
Kate E
KE
Pennick
D000818:Animals; D001921:Brain / Q000821:virology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D007668:Kidney / Q000821:virology; D008168:Lung / Q000821:virology; D010252:Paramyxoviridae / Q000235:genetics / Q000302:isolation & purification*; D018184:Paramyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012911:Snakes / Q000821:virology*
D018076:DNA, Complementary; D012367:RNA, Viral