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LETTER TO THE EDITOR

Dear Sir:

In a recent issue of the American Journal of Tropical Medicine and Hygiene, Johansen and others¹ reported intriguing data on preservation of Japanese encephalitis virus RNA in dead mosquito tissues. Apart from silica gel and/or thymol used for preservation of viral RNA, it might be worth using nucleic acid preservatives. For example, incorporation of a commercially available RNA preservative (RNAlater^®; Ambion, Austin, TX) and an alcohol-based fixative and DNA preservative (GenoFix; DNA Genotek, Inc., Ottawa, Ontario, Canada) might ensure viral viability and RNA stability even under adverse environmental conditions.

Well-preserved host and viral nucleic acids in pigs or mosquitoes in any sentinel system for Japanese encephalitis virus¹ or other vector-borne diseases would effectively address the changing global climate. Moreover, individual ingredients used in molecular biology studies, including any reverse transcriptase or seminested polymerase chain (PCR) technology, have to be constantly stored frozen either at sub-zero temperatures or maintained at 2–8°C. Any inadvertent exposure to temperatures outside the stipulated range would alter their stability, which would lead, in all probability, to erroneous results. Such unfortunate events should not be unexpected anywhere: the massive power cut in California in 2001 could always be repeated!

Surely, integrated investigations that address stability of the ingredients used in PCRs and with nucleic acids would offer an ideal protocol for detection of flaviruses during surveillance in enzootic foci in northern Australia¹ or elsewhere.

Acknowledgment: The secretarial assistance of Seema George is acknowledged.

1. Johansen CA, Hall RA, van den Hurk AF, Ritchie SA, Mackenzie JS, 2002. Detection and stability of Japanese encephalitis virus RNA and virus viability in dead infected mosquitoes under different storage conditions. Am J Trop Med Hyg 67: 656–661.[Abstract]

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