Document 0290
 DOCN  M9460290
 TI    Design and use of signature primers to detect carry-over of amplified
       material.
 DT    9408
 AU    Abbott LZ; Spicer T; Bryz-Gornia V; Kwok S; Sninsky J; Poiesz B;
       Department of Medicine, SUNY Health Science Center, Syracuse; 13210.
 SO    J Virol Methods. 1994 Jan;46(1):51-9. Unique Identifier : AIDSLINE
       MED/94230651
 AB    Signature primer pairs designed for use with the polymerase chain
       reaction have been developed which can determine if a positive result
       originated from the intended target nucleic acid or from so-called
       carry-over contamination of previously amplified DNA. The 3' ends of
       each signature primer, SK339/341, SSK110/111, and SSK58/59 contain a
       viral specific sequence complementary to regions of either HIV-1, HTLV-I
       and II respectively. The 5' ends of each primer contain a non-human,
       non-viral (NHNV) signature sequence including restriction endonuclease
       sites for subsequent cloning. A fourth set of primers, SK338/340,
       consist solely of these NHNV sequences and are designed to anneal to any
       product previously amplified by the viral-specific signature primers.
       These primers were tested against their corresponding positive and
       negative DNA targets, to determine their specificity and sensitivity. As
       expected, the viral-specific signature primers detected the retroviral
       infected samples while no detectable amplification occurred in negative
       DNA controls. Primers SK338/340 did not amplify any viral positive or
       negative template DNA's. Samples spiked with amplified material
       generated from the viral-specific signature primers could be
       specifically amplified by the NHNV primers SK338/340. Primers SK338/340
       were determined to be more sensitive than the viral-specific signature
       primers, ensuring the detection of extremely low amounts of carryover.
       This strategy may be useful in developing other retroviral or
       non-retroviral primers with a built-in signature sequence that can
       differentiate false positives from true positives in a subsequent
       confirmatory test.
 DE    *Artifacts  Base Sequence  Cell Line  Comparative Study  *DNA Primers
       DNA, Viral/*ISOLATION & PURIF  Equipment Contamination  False Positive
       Reactions  HIV-1/GENETICS/*ISOLATION & PURIF  HTLV-I/GENETICS/*ISOLATION
       & PURIF  HTLV-II/GENETICS/*ISOLATION & PURIF  Molecular Sequence Data
       Polymerase Chain Reaction/*METHODS  Proviruses/GENETICS/*ISOLATION &
       PURIF  Sensitivity and Specificity  Support, U.S. Gov't, P.H.S.  JOURNAL
       ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).