SIMPLIFIED REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION PROCEDURE WITH DETECTION BY MICROPLATE HYBRIDIZATION FOR ROUTINE SCREENING OF HEPATITIS-A VIRUS

Reverse transcription polymerase chain reaction, using either nested o r seminested primers, is used extensively for the detection of viruses in small quantities. However, existing methods are prone to false pos itive reactions. We report here an improved polymerase chain reaction technique based on the use of longer primers (39 nucleotides) with sin gle-step amplification, applied to the detection of hepatitis A in low quantities. While the sensitivity of this technique (10 x the 50% tis sue culture infective dose) is equivalent to that of existing methods, it is a simpler procedure, less time consuming, and less susceptible to contamination and therefore provides a more reliable tool for routi ne diagnosis. Finally, the development of a DNA enzyme immunoassay det ection technique and the complete automation of the procedure allow a large number of samples to be processed in clinical laboratories.