1887

Abstract

Summary

Fifteen randomly selected nasopharyngeal (NP) swab specimens (culture-negative for influenza A virus) were spiked with influenza A virus and the nucleic acids were extracted and subjected to PCR amplification with () and (Tth) DNA polymerases. Products of the expected size, and giving equivalent band intensities, were obtained from four specimens with both polymerases. Fox six specimens, less products were obtained with DNA polymerase than with DNA polymerase. Products were detected from five NPs only by PCR with DNA polymerase. the transport medium and the calcium alginate swab fibre of the specimens were shown not to be the source of the inhibitors. the incorporation of P-dCTP into cDNA, and the yield of PCR products of cDNA made from control RNA template (purified from HO spiked virus suspension) were decreased in the presence of inhibitory extracts, showing that both the reverse transcription (RT) and PCR steps in amplification with DNA polymerase were sensitive to the inhibitors. in contrast, DNA polymerase was more resistant to the inhibitors and viral nucleic acid from all the specimens examined could be amplified and detected in a single step by RT-PCR with DNA polymerase.

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/content/journal/jmm/10.1099/00222615-47-12-1131
1998-12-01
2022-08-19
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