Abstract
Abstract: :
Purpose:Epidemics of acute adenovirus conjunctivitis occur in cycles of every 3–4 years in Chennai,India.Polymerase chain reaction–based restriction fragment length polymorphism has been used to rapidly identify a few specific serotypes.1 Since different serotypes were involved in each epidemic and type–specific antisera are not available, a multiplex PCR (mPCR) 2 was standardized and applied to rapidly identify the subgenus group of the causative adenovirus. Methods:During the epidemic in August – December 2002, 85 conjunctival swabs from 62 patients with acute conjunctivitis were tested for direct adenovirus antigen detection by immunofluorescence (IF) and culture using Vero cell line. The extracted DNA from the clinical specimen was amplified using Fastype conjunctivitis adenovirus SSP detection kit (Alcon). The isolated virus was typed by multiplex PCR using subgenus specific primers for subgenera B, C and E (serotypes of these groups are common in this region) targeted against the hexon gene of adenovirus. Results:Of the 85 conjunctival swabs processed, 77 (90.0%) specimens were positive for Adenovirus specific gene by PCR and of these 63 (74.1%) were positive by IF staining. Adenoviruses isolated from10 (11.8%) of these specimens were identified as subgenus E (serotype 4) by mPCR. Neutralization test using adenovirus serotype 4 specific antiserum confirmed the result. Conclusion: mPCR is a rapid, sensitive and specific technique to identify the subgenus group of adenovirus isolates. 1. Madhavan H.N, Therese KL, Dalapathy S, Biswas J. ARVO abstract, IOVS, 39:S435, 1998. 2. Pring –Akerblom, P, Trijseenaar FC, Adrian T, Hoyer H. J Med Virol. 58:87–92, 1999.
Keywords: adenovirus • conjunctivitis • clinical laboratory testing