DNA sequencing validation of Chlamydia trachomatis and Neisseria gonorrhoeae nucleic acid tests.

Abstract:

:DNA sequencing was used to confirm Chlamydia trachomatis and Neisseria gonorrhoeae nucleic acids in endocervical swab samples. DNA in residues of the samples with positive results by 2 commercial kits was subjected to nested polymerase chain reaction (PCR) amplification. The nested PCR amplicons were used as templates for direct automated DNA sequencing. A 40-base signature sequence was sufficient to achieve unequivocal validation of C trachomatis cryptic plasmid and gonococcal opa gene DNA. DNA with a signature sequence specific for C trachomatis was identified in all 14 samples and for N gonorrhoeae in all 13 samples with positive results by the commercial kits for these 2 microbes. In a low-prevalence population, PCR retesting of 289 samples with initial negative results by a non-nucleic acid amplification assay revealed 3 samples positive for C trachomatis and 2 samples positive for N gonorrhoeae that were missed by the commercial kit. DNA sequencing is a useful tool in validating molecular diagnostics.

journal_name

Am J Clin Pathol

authors

Lee SH,Vigliotti VS,Pappu S

doi

10.1309/Y2202RVCGRWW1RWB

subject

Has Abstract

pub_date

2008-06-01 00:00:00

pages

852-9

issue

6

eissn

0002-9173

issn

1943-7722

pii

G36616512913NHU7

journal_volume

129

pub_type

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