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Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine.
Sex Transm Infect. 2010 Jun; 86(3):207-11.ST

Abstract

OBJECTIVE

To evaluate a sensitive and specific, real-time PCR assay with internal control for Chlamydia trachomatis and Neisseria gonorrhoeae DNA detection in urine specimens.

METHODS

The diagnostic performance of a laboratory-developed quadruplex assay (LDQA) targeting the cryptic plasmid and MOMP genes of C trachomatis, the porA pseudogene of N gonorrhoeae and a synthetic internal control was assessed using 1028 urine specimens. The LDQA was compared with the Roche COBAS Taqman CT test and the COBAS Amplicor NG assay with supplemental confirmation tests. The subsequent performance of the LDQA in detecting N gonorrhoeae was monitored in comparison with bacterial culture from swabs.

RESULTS

88 (8.6%) urines were determined as C trachomatis positive in the diagnostic evaluation. LDQA sensitivity and specificity were calculated to be 100% and 99.9%, respectively, for C trachomatis. The LDQA showed high specificity with isolates of other Neisseria species and gave complete concordance with resolved data for N gonorrhoeae detection. However, the incidence of N gonorrhoeae infection was low, with 17 (1.7%) positive patients. A post-implementation audit of 14 316 patients gave the LDQA N gonorrhoeae urine PCR protocol (porA, OPA, 16s rDNA) a sensitivity of 96.9% and specificity of 99.8% in comparison with bacterial culture from swabs.

CONCLUSIONS

The LDQA was found to be an effective method for the detection of C trachomatis and N gonorrhoeae DNA in urine samples, and the PCR protocol has replaced bacterial culture for the screening of N gonorrhoeae in asymptomatic men and women in the laboratory.

Authors+Show Affiliations

Liverpool Specialist Virology Centre, Royal Liverpool University Hospital, Liverpool, UK. m.hopkins@liv.ac.ukNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't
Validation Study

Language

eng

PubMed ID

20522633

Citation

Hopkins, M J., et al. "Validation of a Laboratory-developed Real-time PCR Protocol for Detection of Chlamydia Trachomatis and Neisseria Gonorrhoeae in Urine." Sexually Transmitted Infections, vol. 86, no. 3, 2010, pp. 207-11.
Hopkins MJ, Ashton LJ, Alloba F, et al. Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine. Sex Transm Infect. 2010;86(3):207-11.
Hopkins, M. J., Ashton, L. J., Alloba, F., Alawattegama, A., & Hart, I. J. (2010). Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine. Sexually Transmitted Infections, 86(3), 207-11. https://doi.org/10.1136/sti.2009.040634
Hopkins MJ, et al. Validation of a Laboratory-developed Real-time PCR Protocol for Detection of Chlamydia Trachomatis and Neisseria Gonorrhoeae in Urine. Sex Transm Infect. 2010;86(3):207-11. PubMed PMID: 20522633.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine. AU - Hopkins,M J, AU - Ashton,L J, AU - Alloba,F, AU - Alawattegama,A, AU - Hart,I J, PY - 2010/6/5/entrez PY - 2010/6/5/pubmed PY - 2010/7/29/medline SP - 207 EP - 11 JF - Sexually transmitted infections JO - Sex Transm Infect VL - 86 IS - 3 N2 - OBJECTIVE: To evaluate a sensitive and specific, real-time PCR assay with internal control for Chlamydia trachomatis and Neisseria gonorrhoeae DNA detection in urine specimens. METHODS: The diagnostic performance of a laboratory-developed quadruplex assay (LDQA) targeting the cryptic plasmid and MOMP genes of C trachomatis, the porA pseudogene of N gonorrhoeae and a synthetic internal control was assessed using 1028 urine specimens. The LDQA was compared with the Roche COBAS Taqman CT test and the COBAS Amplicor NG assay with supplemental confirmation tests. The subsequent performance of the LDQA in detecting N gonorrhoeae was monitored in comparison with bacterial culture from swabs. RESULTS: 88 (8.6%) urines were determined as C trachomatis positive in the diagnostic evaluation. LDQA sensitivity and specificity were calculated to be 100% and 99.9%, respectively, for C trachomatis. The LDQA showed high specificity with isolates of other Neisseria species and gave complete concordance with resolved data for N gonorrhoeae detection. However, the incidence of N gonorrhoeae infection was low, with 17 (1.7%) positive patients. A post-implementation audit of 14 316 patients gave the LDQA N gonorrhoeae urine PCR protocol (porA, OPA, 16s rDNA) a sensitivity of 96.9% and specificity of 99.8% in comparison with bacterial culture from swabs. CONCLUSIONS: The LDQA was found to be an effective method for the detection of C trachomatis and N gonorrhoeae DNA in urine samples, and the PCR protocol has replaced bacterial culture for the screening of N gonorrhoeae in asymptomatic men and women in the laboratory. SN - 1472-3263 UR - https://www.unboundmedicine.com/medline/citation/20522633/Validation_of_a_laboratory_developed_real_time_PCR_protocol_for_detection_of_Chlamydia_trachomatis_and_Neisseria_gonorrhoeae_in_urine_ L2 - http://sti.bmj.com/cgi/pmidlookup?view=long&pmid=20522633 DB - PRIME DP - Unbound Medicine ER -