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Nested polymerase chain reaction for detection of human immunodeficiency virus type 1 DNA in clinical specimens

✍ Scribed by Maurizio Zazzi; Laura Romano; Agostina Brasini; Pier Egisto Valensin

Publisher
John Wiley and Sons
Year
1992
Tongue
English
Weight
345 KB
Volume
38
Category
Article
ISSN
0146-6615

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✦ Synopsis

Abstract

A highly sensitive two‐step polymerase chain reaction (PCR) method was evaluated for detection of human immunodeficiency virus type 1 (HIV‐1) DNA in clinical specimens. The product resulting from the first amplification reaction is used as the template for the second PCR with an internal (nested) primer pair. Even when starting from a single copy of HIV‐1 DNA, the double PCR product was readily detected by direct visualization in ethidium bromide‐stained agarose gels. Amplification of minute amounts of HIV‐1 DNA was successful in a considerable excess of HIV‐1 negative DNA than reported previously. All of 85 HIV‐1‐infected individuals were PCR‐positive with at least two of the three sets of primers used, 252 of 255 amplifications allowing unambiguous visualization of a unique DNA fragment of the expected size. The two‐step amplification protocol is simple and rapid and fulfills the requirements of sensitivity and specificity for use in a clinical laboratory. © 1992 Wiley‐Liss, Inc.

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