TY - JOUR
T1 - Evaluation of a new multiplex polymerase chain reaction assay STDFinder for the simultaneous detection of 7 sexually transmitted disease pathogens
AU - Muvunyi, Claude Mambo
AU - Dhont, Nathalie
AU - Verhelst, Rita
AU - Crucitti, Tania
AU - Reijans, Martin
AU - Mulders, Brit
AU - Simons, Guus
AU - Temmerman, Marleen
AU - Claeys, Geert
AU - Padalko, Elizaveta
N1 - Funding Information:
The authors would like to thank the staff at the Department of Clinical Chemistry, Microbiology and Immunology, Center for Molecular Diagnostics, Ghent University Hospital for their technical assistance. They also thank Yvonne Hong from Emory University, Atlanta, GA, for her careful English editing of the manuscript. This study was approved by the National Ethics Committee of Rwanda and the University of Ghent Ethics Committee in Ghent, Belgium, and financially supported by a PhD grant from Ghent University, Ghent, Belgium, through its VLIR (Flemish Interuniversity Council) Own Initiative (project number VLIR-UOS ZEIN2007PR342-19878 ).
PY - 2011/9
Y1 - 2011/9
N2 - We evaluated a new multiplex polymerase chain reaction (mPCR), "STDFinder assay", a novel multiplex ligation-dependent probe amplification (MLPA) assay for the simultaneous detection of 7 clinically relevant pathogens of STDs, i.e., Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Mycoplasma genitalium, Treponema pallidum, and herpes simplex virus type 1 and 2 (HSV-1 and HSV-2). An internal amplification control was included in the mPCR reaction. The limits of detection for the STDFinder assay varied among the 7 target organisms from 1 to 20 copies per MLPA assay. There were no cross-reactions among any of the probes. Two hundred and forty-two vaginal swabs and an additional 80 specimens with known results for N. gonorrhoeae and C. trachomatis, obtained from infertile women seen at an infertility research clinic at the Kigali Teaching Hospital in Rwanda, were tested by STDFinder assay and the results were confirmed by single real-time PCR using different species-specific targets. Compared to the reference standard, the STDFinder assay showed specificities and sensitivities of 100% and 100%, respectively, for N. gonorrhoeae, C. trachomatis, and M. genitalium; 90.2% and 100%, respectively, for Trichomonas vaginalis; and 96.1% and 100%, respectively, for HSV-2. No specimen was found to be positive for HSV-1 by either the STDFinder assay or the comparator method. Similarly, the sensitivity for Treponema pallidum could not be calculated due to the absence of any Treponema pallidum-positive samples. In conclusion, the STDFinder assays have comparable clinical sensitivity to the conventional mono and duplex real-time PCR assay and are suitable for the routine detection of a broad spectrum of these STDs at relatively low cost due to multiplexing.
AB - We evaluated a new multiplex polymerase chain reaction (mPCR), "STDFinder assay", a novel multiplex ligation-dependent probe amplification (MLPA) assay for the simultaneous detection of 7 clinically relevant pathogens of STDs, i.e., Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Mycoplasma genitalium, Treponema pallidum, and herpes simplex virus type 1 and 2 (HSV-1 and HSV-2). An internal amplification control was included in the mPCR reaction. The limits of detection for the STDFinder assay varied among the 7 target organisms from 1 to 20 copies per MLPA assay. There were no cross-reactions among any of the probes. Two hundred and forty-two vaginal swabs and an additional 80 specimens with known results for N. gonorrhoeae and C. trachomatis, obtained from infertile women seen at an infertility research clinic at the Kigali Teaching Hospital in Rwanda, were tested by STDFinder assay and the results were confirmed by single real-time PCR using different species-specific targets. Compared to the reference standard, the STDFinder assay showed specificities and sensitivities of 100% and 100%, respectively, for N. gonorrhoeae, C. trachomatis, and M. genitalium; 90.2% and 100%, respectively, for Trichomonas vaginalis; and 96.1% and 100%, respectively, for HSV-2. No specimen was found to be positive for HSV-1 by either the STDFinder assay or the comparator method. Similarly, the sensitivity for Treponema pallidum could not be calculated due to the absence of any Treponema pallidum-positive samples. In conclusion, the STDFinder assays have comparable clinical sensitivity to the conventional mono and duplex real-time PCR assay and are suitable for the routine detection of a broad spectrum of these STDs at relatively low cost due to multiplexing.
KW - Chlamydia trachomatis
KW - Herpes simplex virus type 1/2
KW - Multiplex PCR
KW - Mycoplasma genitalium
KW - Neisseria gonorrhoeae
KW - STDFinder
KW - Treponema pallidum
KW - Trichomonas vaginalis
UR - http://www.scopus.com/inward/record.url?scp=80051663818&partnerID=8YFLogxK
U2 - 10.1016/j.diagmicrobio.2011.06.005
DO - 10.1016/j.diagmicrobio.2011.06.005
M3 - Article
C2 - 21798683
AN - SCOPUS:80051663818
SN - 0732-8893
VL - 71
SP - 29
EP - 37
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 1
ER -
