A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the M-PCR assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results were available for 99 specimens collected during the third interval. Confirmatory PCR assays targeting different gene sequences for each of the three organisms were used to validate the M-PCR results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M-PCR and the confirmatory PCR were positive. The resolved sensitivities of M-PCR for HSV, H. ducreyi, and T. pallidum were 100, 98.4, and 91\%, respectively. The resolved sensitivities of HSV culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81\%, respectively. These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.
%0 Journal Article
%1 orle_simultaneous_1996
%A Orle, K A
%A Gates, C A
%A Martin, D H
%A Body, B A
%A Weiss, J B
%D 1996
%J Journal of Clinical Microbiology
%K 1, 2, Bacterial, Bacteriological Base Chain Chancroid, Colorimetry, Data, Diseases, Genital Genitalis, Haemophilus Herpes Herpesvirus Human, Humans, Male, Molecular Polymerase Primers, Probes, Reaction, Sensitivity Sequence Sequence, Specificity, Syphilis, Techniques, Treponema Ulcer, Viral, Virology and ducreyi, pallidum, {DNA,} {DNA}
%N 1
%P 49--54
%T Simultaneous PCR detection of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus types 1 and 2 from genital ulcers
%U http://www.ncbi.nlm.nih.gov/pubmed/8748271
%V 34
%X A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the M-PCR assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results were available for 99 specimens collected during the third interval. Confirmatory PCR assays targeting different gene sequences for each of the three organisms were used to validate the M-PCR results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M-PCR and the confirmatory PCR were positive. The resolved sensitivities of M-PCR for HSV, H. ducreyi, and T. pallidum were 100, 98.4, and 91\%, respectively. The resolved sensitivities of HSV culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81\%, respectively. These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.
@article{orle_simultaneous_1996,
abstract = {A multiplex {PCR} {(M-PCR)} assay with colorimetric detection was devised for the simultaneous amplification of {DNA} targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus {(HSV)} types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the {M-PCR} assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. {HSV} culture results were available for 99 specimens collected during the third interval. Confirmatory {PCR} assays targeting different gene sequences for each of the three organisms were used to validate the {M-PCR} results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the {M-PCR} and the confirmatory {PCR} were positive. The resolved sensitivities of {M-PCR} for {HSV,} H. ducreyi, and T. pallidum were 100, 98.4, and 91\%, respectively. The resolved sensitivities of {HSV} culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81\%, respectively. These results indicate that the {M-PCR} assay is more sensitive than standard diagnostic tests for the detection of {HSV,} H. ducreyi, and T. pallidum from genital ulcers.},
added-at = {2011-03-11T10:05:34.000+0100},
author = {Orle, K A and Gates, C A and Martin, D H and Body, B A and Weiss, J B},
biburl = {https://www.bibsonomy.org/bibtex/26c19b62256f32dea0ea89edc77d04ea4/jelias},
interhash = {d3e0109212cb2e58ec8b0f2bc3fc9fba},
intrahash = {6c19b62256f32dea0ea89edc77d04ea4},
issn = {0095-1137},
journal = {Journal of Clinical Microbiology},
keywords = {1, 2, Bacterial, Bacteriological Base Chain Chancroid, Colorimetry, Data, Diseases, Genital Genitalis, Haemophilus Herpes Herpesvirus Human, Humans, Male, Molecular Polymerase Primers, Probes, Reaction, Sensitivity Sequence Sequence, Specificity, Syphilis, Techniques, Treponema Ulcer, Viral, Virology and ducreyi, pallidum, {DNA,} {DNA}},
month = jan,
note = {{PMID:} 8748271},
number = 1,
pages = {49--54},
timestamp = {2011-03-11T10:06:21.000+0100},
title = {Simultaneous {PCR} detection of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus types 1 and 2 from genital ulcers},
url = {http://www.ncbi.nlm.nih.gov/pubmed/8748271},
volume = 34,
year = 1996
}