Detection of HSV mRNA using reverse transcription-polymerase chain reaction for diagnosis in murine herpetic keratitis model.
Reverse transcription polymerase chain reaction (RT-PCR) was applied in the detection of herpes simplex virus-1 (HSV-1) mRNA from tear film and corneal epithelium in a murine herpetic keratitis model. The diagnostic value of this new technique for acute herpetic keratitis was evaluated in comparison with direct PCR for genomic DNA and viral culture. On day 2 postinfection (PI) of HSV, all mice showed dendritic keratitis, and PCR, RT-PCR, and viral culture were positive in all samples. On day 8 PI, no dendritic keratitis was observed in any mouse, PCR was positive in all samples, while RT-PCR was positive in only 5 of 12 samples and viral culture in only 2 of 12. The sensitivity of RT-PCR was lower than that of PCR, and approximately the same as viral culture; however, the findings of RT-PCR more closely concurred with clinical observations than the findings of PCR. These results show the potential of RT-PCR for rapid, specific diagnosis of acute herpetic keratitis.
Mori
Y
Y
Department of Microbiology, Osaka University Medical School, Japan.
Inoue
Y
Y
Shimomura
Y
Y
Kase
T
T
Tano
Y
Y
D000208:Acute Disease; D000818:Animals; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000821:virology*; D005260:Female; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000302:isolation & purification; D016849:Keratitis, Herpetic / Q000175:diagnosis*; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000032:analysis*; D012680:Sensitivity and Specificity; D013666:Tears / Q000821:virology*; D014158:Transcription, Genetic; D014776:Virus Cultivation
D012333:RNA, Messenger