Tracing of the molecular remnants of herpes virus infections in necrotic skin tissue.
We have established an assay system to detect herpesvirus-derived transcripts in lesional crusts. Fifteen patients with herpes simplex (HS), 21 with herpes zoster (HZ), 2 with varicella, and 20 with irrelevant diseases were enrolled in the present study. Total RNA was extracted from crusts or scales, and converted to cDNA. Virus-encoded transcripts were amplified using reverse transcriptase (RT)-PCR. Housekeeping gene transcripts such as beta2-microglobulin (beta2-MG) and beta-actin (beta-actin) mRNA were also examined, and an efficient preservative condition of the crusts was determined. With extracted RNAs, beta2-MG and beta-actin mRNA were successfully amplified in all crust samples. Herpes simplex virus (HSV)-specific, lytic cycle-related transcript, UL30 mRNA was detected in all 15 HS samples, including 13 samples of HSV-1- and 2 of HSV-2-encoded UL30 mRNA, respectively. Of 23 samples, including 21 HZ and 2 varicella cases, varicella zoster virus (VZV)-specific, lytic cycle-related transcript, ORF40 mRNA was detected in 22 samples. In a control group, no UL30 and ORF40 mRNA were detected. Crust samples that had been stored without any pretreatment or preservative for 6 months at room temperature (RT) were available for the present assay. When compared with the freshly obtained materials, the amount of beta2-MG mRNA was reduced to 51% in the stored samples covered with adhesive tape, to 48% in a sample left at R.T. without any treatment, and to 1.2% in the samples stocked in saline for 5 days. Herpes virus- and host-derived transcripts contained in crusts can be detected by RT-PCR amplification. Crusts or dry epidermal necrosis with inflammatory cells may provide beneficial diagnostic information.
Yamamoto
Takenobu
T
Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan. go-yama@md.okayama-u.ac.jp
Yamada
Akiko
A
Tsuji
Kazuhide
K
Iwatsuki
Keiji
K
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D005260:Female; D006561:Herpes Simplex / Q000821:virology*; D006562:Herpes Zoster / Q000821:virology*; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D009336:Necrosis / Q000821:virology; D012367:RNA, Viral / Q000032:analysis*; D018139:Simplexvirus / Q000235:genetics* / Q000302:isolation & purification; D012867:Skin / Q000473:pathology* / Q000821:virology*; D055815:Young Adult
D012367:RNA, Viral