ABSTRACT
Research using nucleic acid amplification tests (NAATs) have repeatedly found rectal and oropharyngeal infections with Chlamydia trachomatis and Neisseria gonorrhoeae to be common and potentially more difficult to treat than genital infections. Unfortunately, public health and patient care efforts have been hampered by the lack of FDA-cleared NAATs with claims for anorectal or oropharyngeal samples. At the time of the initiation of this study, no commercially available assays had these claims. We formed a novel partnership among academic institutions and diagnostic manufacturers to address this public health need. From May 2018 through August 2019, we recruited 1108 women, 1256 men, and 26 transgender persons each of whom provided 3 anal and 3 oropharyngeal swab specimens. The 3 anal swabs were pooled into a single transport tube as were the 3 oropharyngeal swabs. The performance of each of three study assays was estimated by comparison to the composite result and relative to one another. Percent positivity for chlamydia was 5.9 and 1.2% from anal and oropharyngeal specimens, respectively, compared to 4.2 and 4.1% for gonorrhea. Sensitivity for chlamydia detection ranged from 81.0 to 95.1% and 82.8 to 100% for anal and oropharyngeal specimens, respectively. Gonorrhea sensitivity ranged from 85.9 to 99.0% and 74.0 to 100% for anal and oropharyngeal samples, respectively. Specificity estimates were ≥ 98.9% for all assays, organisms, and sample types. Although there was heterogeneity between sensitivity estimates, these assays offer better ability to detect extragenital infections than culture and potential solutions for providing appropriate sexual health care for populations in which these infections are of concern.
Subject(s)
Chlamydia Infections , Gonorrhea , Chlamydia Infections/diagnosis , Chlamydia trachomatis/genetics , Female , Gonorrhea/diagnosis , Humans , Male , Neisseria gonorrhoeae/genetics , Nucleic Acid Amplification Techniques , Sensitivity and SpecificityABSTRACT
Chlamydia trachomatis is the world's most prevalent bacterial sexually transmitted infection and leading infectious cause of blindness, yet it is one of the least understood human pathogens, in part due to the difficulties of in vitro culturing and the lack of available tools for genetic manipulation. Genome sequencing has reinvigorated this field, shedding light on the contemporary history of this pathogen. Here, we analyze 563 full genomes, 455 of which are novel, to show that the history of the species comprises two phases, and conclude that the currently circulating lineages are the result of evolution in different genomic ecotypes. Temporal analysis indicates these lineages have recently expanded in the space of thousands of years, rather than the millions of years as previously thought, a finding that dramatically changes our understanding of this pathogen's history. Finally, at a time when almost every pathogen is becoming increasingly resistant to antimicrobials, we show that there is no evidence of circulating genomic resistance in C. trachomatis.
Subject(s)
Chlamydia trachomatis/genetics , Drug Resistance, Bacterial/genetics , Ecotype , Evolution, Molecular , Genome, Bacterial , Chlamydia trachomatis/isolation & purification , Female , Humans , MaleABSTRACT
Evaluating the clinical performance of a new nucleic acid amplification test (NAAT) for Mycoplasma genitalium, B. Kirkconnell, B. Weinbaum, K. Santos, T. Le Nguyen, et al. (J Clin Microbiol 57:e00264-19, 2019, https://doi.org/10.1128/JCM.00264-19) created 3 alternate NAATs that detected other unique M. genitalium gene targets. Lacking a reference standard, they used the consensus of results with those 3 NAATs as the comparator. This approach could be a new paradigm to evaluate new NAATs when there is no previously defined reference standard.
Subject(s)
Mycoplasma genitalium/genetics , Nucleic Acids , Nucleic Acid Amplification TechniquesABSTRACT
Self-obtained vaginal swabs, first-void urine and pooled specimens were collected at home and in a clinic. Percent prevalence and collection site concordance was 30.3 and 100 for Mycoplasma genitalium (74.4% azithromycin resistant) 15.1 and 96.7 for Chlamydia trachomatis and 6.6 and 100 for Neisseria gonorrhoeae (27% ciprofloxacin-resistant).
Subject(s)
Chlamydia Infections/diagnosis , Chlamydia trachomatis/genetics , Gonorrhea/diagnosis , Mycoplasma Infections/diagnosis , Mycoplasma genitalium/genetics , Neisseria gonorrhoeae/genetics , Vagina/microbiology , Adolescent , Adult , Ambulatory Care Facilities/statistics & numerical data , Chlamydia Infections/urine , Female , Gonorrhea/urine , Humans , Mycoplasma Infections/urine , Nucleic Acid Amplification Techniques , Specimen Handling/methods , Young AdultABSTRACT
The Aptima Combo 2 (AC2) and Aptima CT (ACT) (Hologic Inc., San Diego, CA) are nucleic acid amplification tests (NAATs) that detect Chlamydia trachomatis AC2 also detects Neisseria gonorrhoeae Storage and temperature conditions may impact the utility of NAATs in some settings and screening programs. We evaluated specimen stability for use beyond the Aptima package insert specifications for temperature and duration of storage (between 2°C and 30°C and 60 days, respectively) in two studies: (i) dry C. trachomatis-seeded swabs were used with ACT after storage at 4°C, 23°C, or 36°C for up to 84 days and (ii) swabs seeded with C. trachomatis and N. gonorrhoeae and then placed in transport medium were tested with AC2, after being mailed via the U.S. Postal Service to three different sites. Prolonged storage of samples had no effect, and samples stored at 4°C, 23°C, and 36°C for up to 84 days yielded comparable ACT positivities, although there was a drop in signal intensity for virtually all specimens under all storage/shipping conditions after day 21. In the mailing study, 80%, 52% and 29% of seeded swabs were exposed to temperatures of >30°C during three rounds in transit, and 2% reached temperatures of >40°C. No evidence of signal degradation in the AC2 assay for detection of C. trachomatis or N. gonorrhoeae was observed, although some mailed swabs took more than 5 weeks to reach the laboratory site. These two studies support the potential use of swabs at temperatures above 36°C and storage beyond 60 days and provide confidence regarding this commercially available NAAT for testing of specimens after mailing.
Subject(s)
Bacteriological Techniques/methods , Chlamydia Infections/diagnosis , Chlamydia trachomatis/isolation & purification , Gonorrhea/diagnosis , Neisseria gonorrhoeae/isolation & purification , Specimen Handling/methods , Humans , Temperature , Time Factors , United StatesABSTRACT
We evaluated Abbott's RealTime assay for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) in the urethra, oropharynx, and rectum of 260 men who have sex with men. Compared with Hologic's AC2, RealTime had good agreement for detecting CT and GC. Overall, there were 25 CT and 44 GC AC2 positives, and 26 CT and 38 GC RealTime positives. For total negatives, there were 742 CT and 725 GC for AC2, 744 CT and 724 GC for RealTime.
Subject(s)
Bacterial Typing Techniques/instrumentation , Chlamydia Infections/diagnosis , Chlamydia trachomatis/isolation & purification , Gonorrhea/diagnosis , Homosexuality, Male , Neisseria gonorrhoeae/isolation & purification , Oropharynx/microbiology , Real-Time Polymerase Chain Reaction/instrumentation , Rectum/microbiology , Urethra/microbiology , Adult , Chlamydia Infections/prevention & control , Chlamydia Infections/transmission , Chlamydia trachomatis/genetics , Gonorrhea/prevention & control , Gonorrhea/transmission , Humans , Male , Neisseria gonorrhoeae/genetics , Reagent Kits, Diagnostic , San Francisco/epidemiology , Sensitivity and SpecificityABSTRACT
BACKGROUND: Anorectal and pharyngeal infections with Chlamydia trachomatis (CT) and Neisseria gonorrheae (NG) are commonly observed in men who have sex with men (MSM). There is increasing evidence that such infections at extra-genital sites are also common in women. In both sexes, these infections are largely overlooked as they are not routinely tested for in regular care. Testing based on sexual behavior or symptoms would only detect half of these extra-genital infections. This paper elucidates the differences and similarities between women and MSM, regarding the epidemiology of extra-genital CT and NG. It discusses the clinical and public health impact of untested extra-genital infections, how this may impact management strategies, and thereby identifies key research areas. DISCUSSION: Extra-genital CT is as common in women as it is in MSM; NG in women is as common at their extra-genital sites as it is at their genital sites. The substantial numbers of extra-genital CT and NG being missed in women and MSM indicate a need to test and treat more patients and perhaps different choices in treatment and partner management strategies. Doing so will likely contribute to reduced morbidity and transmission in both sexes. However, in our opinion, it is clear that there are several knowledge gaps in understanding the clinical and public health impact of extra-genital CT and NG. Key research areas that need to be addressed concern associated morbidity (anorectal and reproductive morbidity due to extra-genital infections), 'the best' management strategies, including testing and treatment for extra-genital CT, extra-genital treatment resistance, transmission probabilities between partners and between anatomic sites in a woman, and impact on transmission of other infections. Data are also lacking on cost-effectiveness of pharyngeal testing, and of NG testing and anorectal CT testing in women. Gaps in the management of extra-genital CT and NG may also apply for other STIs, such Mycoplasma genitalium. Current management strategies, including testing, to address extra-genital CT and NG in both sexes are suboptimal. Comparative data on several identified key themes in women and MSM are lacking and urgently needed to guide better management of extra-genital infections.
Subject(s)
Chlamydia Infections/epidemiology , Chlamydia trachomatis/pathogenicity , Gonorrhea/epidemiology , Neisseria gonorrhoeae/pathogenicity , Pharyngeal Diseases/microbiology , Rectal Diseases/microbiology , Chlamydia Infections/diagnosis , Chlamydia Infections/drug therapy , Cost-Benefit Analysis , Female , Gonorrhea/diagnosis , Gonorrhea/drug therapy , Guidelines as Topic , Homosexuality, Male , Humans , Male , Mycoplasma genitalium/pathogenicity , Pharyngeal Diseases/drug therapy , Pharynx/microbiology , Rectal Diseases/drug therapy , Rectum/microbiology , Sexual Behavior , Sexual PartnersABSTRACT
BACKGROUND: Trachoma, caused by repeated infections with ocular Chlamydia trachomatis, is targeted for elimination using multiple annual rounds of mass drug administration (MDA) in endemic communities. Infection rates do not decline as expected in some communities, leading to concerns about azithromycin resistance. METHODS: After 3 yearly MDAs in 32 communities in Tanzania, 107 children were identified 1 year later with infection. All were provided MDA again, and 90 were seen again at 2 months, of whom 30 had infection. Chlamydia trachomatis isolates were obtained before and after MDA in 15 paired samples and were tested for antimicrobial susceptibility. The infectious load of C. trachomatis before MDA was determined in 30 children who had infection at both times and 60 whose infection cleared. RESULTS: The median load was 8.6 genome copies per polymerase chain reaction in the consistently infected, and 8.4 in those whose infection cleared (P = .86). For the consistently infected, the average minimum inhibitory concentration was 0.26 µg/mL for azithromycin before and 0.20 µg/mL after MDA. All isolates had minimum inhibitory concentration ≤0.50 µg/mL. CONCLUSIONS: There is no evidence that continued infection after MDA was due either to resistance to azithromycin or to a heavier load of organism before treatment. Other potential causes of persistent infection need to be evaluated.
Subject(s)
Anti-Bacterial Agents/administration & dosage , Azithromycin/administration & dosage , Chlamydia trachomatis/drug effects , Drug Resistance, Bacterial , Trachoma/drug therapy , Trachoma/microbiology , Anti-Bacterial Agents/pharmacology , Azithromycin/pharmacology , Child , Child, Preschool , Chlamydia trachomatis/isolation & purification , Drug Therapy/methods , Female , Humans , Infant , Infant, Newborn , Male , Microbial Sensitivity Tests , Secondary Prevention , Tanzania/epidemiology , Trachoma/epidemiology , Trachoma/prevention & controlABSTRACT
A specific real-time polymerase chain reaction followed by melt curve analysis was developed for the detection of the Swedish variant (nvCT) strain of Chlamydia trachomatis (CT). Surveillance was performed on 476 CT-positive clinical specimens obtained from 15 laboratories around the United States using nucleic acid amplification test assays, which would not miss the nvCT. All were negative for nvCT; thus, there is no evidence of the nvCT in the United States.
Subject(s)
Chlamydia Infections/epidemiology , Chlamydia trachomatis/isolation & purification , Communicable Diseases, Emerging/epidemiology , Chlamydia Infections/classification , Chlamydia Infections/genetics , Chlamydia trachomatis/classification , Communicable Diseases, Emerging/genetics , Female , Genotype , Humans , Male , Norway/epidemiology , Nucleic Acid Amplification Techniques , Prevalence , Real-Time Polymerase Chain Reaction , Sexual Behavior , Sweden/epidemiology , United States/epidemiologyABSTRACT
Twelve trachoma-hyperendemic communities were treated with 3 annual mass azithromycin distributions. Children aged 0-9 years were monitored 1 year following the third treatment. An RNA-based test detected ocular chlamydial infection in more children than did a DNA-based test (6.9% vs 4.2%), and in a larger number of communities (8 vs 7).
Subject(s)
Anti-Bacterial Agents/administration & dosage , Azithromycin/administration & dosage , Chlamydia trachomatis/isolation & purification , RNA, Bacterial/analysis , RNA, Ribosomal/analysis , Trachoma/prevention & control , Antibiotic Prophylaxis/methods , Child , Child, Preschool , Chlamydia trachomatis/genetics , Ethiopia/epidemiology , Female , Humans , Infant , Male , Prevalence , Preventive Health Services , RNA, Bacterial/genetics , RNA, Ribosomal/genetics , Statistics, Nonparametric , Trachoma/drug therapy , Trachoma/epidemiology , Trachoma/microbiologyABSTRACT
We analyzed by multilocus sequence typing 77 lymphogranuloma venereum Chlamydia trachomatis strains from men who have sex with men in Europe and the United States. Specimens from an outbreak in 2003 in Europe were monoclonal. In contrast, several strains were in the United States in the 1980s, including a variant from Europe.
Subject(s)
Chlamydia trachomatis/genetics , Lymphogranuloma Venereum/epidemiology , Lymphogranuloma Venereum/microbiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Disease Outbreaks , Europe/epidemiology , Gene Expression Regulation , Genetic Variation , Genotype , Homosexuality, Male , Humans , MaleABSTRACT
Nucleic acid amplification tests are increasingly used to detect ocular chlamydia infection in trachoma research and programs. To evaluate the reliability of Chlamydia trachomatis detection by the Abbott RealTime CT/NG assay (Abbott Molecular, Inc., Des Plaines, IL) on the m2000 platform, three conjunctival samples were collected from each of 200 children aged 0-9 years in Ethiopia: two from the right eye and one from the left eye. Four aliquots were processed for each child: two from the first right eye sample, one from the second right eye sample, and one from the left eye sample. Sixty-nine swabs were processed in a U.S. laboratory and 131 in an Ethiopian laboratory. Intra-class correlation coefficients (ICCs) were high when comparing two aliquots from the same swab (ICC ranged from 0.96 to 0.99), two separate swabs from the right eye (0.89-0.91), and one right and one left eye swab (0.87-0.89), indicating reliable chlamydial load assessment across different samples and laboratory settings.
Subject(s)
Chlamydia trachomatis , Conjunctivitis, Inclusion/diagnosis , Nucleic Acid Amplification Techniques/methods , Child , Child, Preschool , Conjunctivitis, Inclusion/epidemiology , Conjunctivitis, Inclusion/microbiology , Cross-Sectional Studies , Ethiopia/epidemiology , Eye/microbiology , Female , Humans , Infant , Infant, Newborn , Male , Sensitivity and SpecificityABSTRACT
BACKGROUND: After approximately 5 years of SAFE (surgery, antibiotics, facial cleanliness, environmental improvement) interventions for trachoma, hyperendemic (trachomatous inflammation-follicular (TF) ≥30%) districts remained in Amhara, Ethiopia. This study's aim was to characterize the epidemiology of Chlamydia trachomatis (Ct) infection and load among pre-school aged children living under the SAFE strategy. METHODS: Conjunctival swabs from a population-based sample of children aged 1-5 years collected between 2011 and 2015 were assayed to provide Ct infection data from 4 endemic zones (comprised of 58 districts). Ct load was determined using a calibration curve. Children were graded for TF and trachomatous inflammation-intense (TI). RESULTS: 7,441 children were swabbed in 4 zones. TF and TI prevalence were 39.9% (95% confidence Interval [CI]: 37.5%, 42.4%), and 9.2% (95% CI: 8.1%, 10.3%) respectively. Ct infection prevalence was 6.0% (95% CI: 5.0%, 7.2%). Infection was highest among children aged 2 to 4 years (6.6%-7.0%). Approximately 10% of infection occurred among children aged 1 year. Ct load decreased with age (P = 0.002), with the highest loads observed in children aged 1 year (P = 0.01) vs. aged 5 years. Participants with TF (P = 0.20) and TI (P<0.01) had loads greater than individuals without active trachoma. CONCLUSIONS: In this hyperendemic setting, it appears that the youngest children may contribute in meaningful ways towards persistent active trachoma.
Subject(s)
Chlamydia trachomatis/physiology , Trachoma/epidemiology , Trachoma/prevention & control , Anti-Bacterial Agents/administration & dosage , Child, Preschool , Chlamydia trachomatis/drug effects , Conjunctiva/microbiology , Endemic Diseases/prevention & control , Ethiopia/epidemiology , Female , Humans , Infant , Male , Trachoma/drug therapy , Trachoma/microbiologyABSTRACT
We investigated antimicrobial drug resistance in ocular Chlamydia trachomatis 18 months after 4 biannual communitywide distributions of antimicrobial drugs in a region of Ethiopia where ocular strains of C. trachomatis are highly endemic. We found no significant differences in susceptibilities to azithromycin and doxycycline in 6 posttreatment and 4 pretreatment samples.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Azithromycin/therapeutic use , Chlamydia trachomatis/drug effects , Drug Resistance, Microbial/genetics , Eye Diseases/microbiology , Administration, Topical , Adult , Child , Child, Preschool , Chlamydia trachomatis/genetics , Cycloheximide/pharmacology , Ethiopia/epidemiology , Eye Diseases/drug therapy , Eye Diseases/epidemiology , Female , Genetic Predisposition to Disease , Humans , Infant , Male , Pregnancy , Pregnancy Complications, Infectious/drug therapy , PrevalenceABSTRACT
Self-collected glans and rectal swab specimens from men who have sex with men (MSM) may be appropriate, convenient specimens for testing. We evaluated the use of self-collected swabs for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by a transcription-mediated amplification test (AC2; Aptima Combo 2; Gen-Probe Inc.) and a strand displacement amplification test (SDA; ProbeTec; Becton Dickinson Co.) in MSM seen at the city sexually transmitted disease clinic in San Francisco, CA. For the glans swab specimen, subjects enrolled early in the study rolled a Dacron swab across the meatus three times (method 1). A slightly more invasive procedure was performed later in the study: the subjects inserted the swab 1/4 in. into the urethra, rotated the swab, and then withdrew the swab (method 2). MSM self-collected a rectal swab specimen and also provided first-catch urine (FCU). Additional rectal swab samples were then obtained by the clinician. For the detection of C. trachomatis and N. gonorrhoeae, all swabs were evaluated by AC2 and SDA, FCU was tested by AC2, and the clinician-collected rectal swabs were cultured. A rectal true-positive (TP) result was defined as a culture-positive result for C. trachomatis or N. gonorrhoeae, two or more positive nucleic acid amplification test (NAAT) results, or a single NAAT-positive result confirmed by an alternate amplification method (the Aptima C. trachomatis or N. gonorrhoeae test). A glans TP result was defined as a positive result for FCU, positive results for both glans specimens (one tested by AC2 and one tested by SDA), or a positive result for a single glans specimen confirmed by an alternate amplification method. The prevalence rates of C. trachomatis and N. gonorrhoeae by testing of FCU were 6.8% (60/882 specimens) and 12.2% (108/882 specimens), respectively. Mixed results were obtained with the glans swab: N. gonorrhoeae detection by AC2 and SDA (method 1) had the best performance (sensitivities, >92%) with samples from a population with a higher prevalence of infection, but their performance for the detection of C. trachomatis was poor and varied by collection method (sensitivities, 56 to 68%). The prevalence rates of C. trachomatis and N. gonorrhoeae in the rectum were 7.3% (66/907 specimens) and 9.4% (83/882 specimens), respectively. The sensitivities of the tests with self-collected and clinician-collected rectal swab specimens were comparable (for C. trachomatis, 41% and 44%, respectively, by SDA and 82% and 71%, respectively, by AC2; for N. gonorrhoeae, 77% and 68%, respectively, by SDA and 84% and 78%, respectively, by AC2). AC2 and SDA were far superior to culture for the detection of C. trachomatis and N. gonorrhoeae in the rectum, with both tests detecting at least twice as many infections. While we found self-collected rectal swabs from MSM to be valid specimens for testing, the sensitivities of the tests with glans swab specimens were disappointing except for those from patients with symptomatic N. gonorrhoeae infections. Self-collected glans swab specimens may not be appropriate for the detection of C. trachomatis or for the detection of N. gonorrhoeae in low-risk or asymptomatic patients by AC2 and SDA, and we would not recommend their use on the basis of our results. Further studies are needed.
Subject(s)
Chlamydia trachomatis/isolation & purification , Neisseria gonorrhoeae/isolation & purification , Nucleic Acid Amplification Techniques/methods , Penis/microbiology , Rectum/microbiology , Self-Examination , Chlamydia Infections/diagnosis , Chlamydia Infections/microbiology , Chlamydia trachomatis/genetics , Gonorrhea/diagnosis , Gonorrhea/microbiology , Health Services Research , Homosexuality, Male , Humans , Male , Neisseria gonorrhoeae/genetics , Prevalence , San Francisco , Sensitivity and SpecificityABSTRACT
OBJECTIVE: To determine whether infectious trachoma can be completely eliminated from severely affected villages. DESIGN: Cross-sectional survey of 2 villages previously enrolled and monitored over 42 months as part of a larger, group-randomized clinical trial. PARTICIPANTS: A total of 758 individuals residing in 2 villages with high baseline trachoma prevalence, of a total population of 768 (98.7%). METHODS: All members of the 2 villages were offered 6 biannual mass treatments with oral azithromycin. At 42 months, each current village member was examined. The right upper tarsal conjunctiva was everted and swabbed. Samples were processed for evidence of Chlamydia trachomatis RNA. MAIN OUTCOME MEASURES: Clinical activity by World Health Organization simplified grading scale for trachoma and laboratory evidence of chlamydial RNA. RESULTS: Average antibiotic coverage over the study period was 90% and 94% in the 2 villages. Clinical trachoma activity in children aged 1 to 5 years decreased from 78% and 83% in the 2 villages before treatment to 17% and 24% at 42 months. Polymerase chain reaction (PCR) evidence of infection in the same age group decreased from 48% to 0% in both villages at 42 months. When all age groups were examined, there were zero cases with evidence of chlamydial RNA among 758 total villagers tested. CONCLUSIONS: Biannual mass distribution of azithromycin can locally eliminate ocular chlamydial infection from severely affected communities.
Subject(s)
Anti-Bacterial Agents/administration & dosage , Azithromycin/administration & dosage , Trachoma/drug therapy , Administration, Oral , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Chlamydia trachomatis/genetics , Chlamydia trachomatis/isolation & purification , Conjunctiva/microbiology , Cross-Sectional Studies , Ethiopia/epidemiology , Female , Humans , Infant , Infant, Newborn , Male , Mass Screening , Middle Aged , Polymerase Chain Reaction , Prevalence , RNA, Bacterial/analysis , Rural Population , Trachoma/epidemiology , Trachoma/microbiology , Young AdultABSTRACT
We evaluated three of the CDC approaches for confirming Neisseria gonorrhoeae (gonococcus [GC])-positive nucleic acid amplification test (NAAT) results: (i) repeating the original test on the original specimen, (ii) testing the original specimen with a different test, and (iii) performing a different test on a duplicate specimen collected at the same visit. For the first approach, clinical specimens were initially tested by Aptima Combo 2 (AC2) (Gen-Probe Inc., San Diego, CA), ProbeTec (strand displacement amplification [SDA]) (Becton Dickinson Co., Sparks, MD), and Amplicor (PCR) (Roche Molecular Systems, Branchburg, NJ). The original GC-positive specimens were then retested by the same NAAT for confirmation. For the second approach, specimens initially positive by AC2, SDA, or PCR were retested by different NAATs (SDA, PCR, AC2, and Aptima Neisseria gonorrhoeae assay [AGC]; Gen-Probe Inc.). For the third approach, duplicate urethral swabs and first-catch urine (FCU) samples from men and duplicate cervical swabs and FCU samples from women were each tested by SDA, AC2, and AGC in parallel. We found that 89 to 96% of samples positive by SDA, PCR, and AC2 were confirmed by repeat testing and that 85 to 98% of SDA, PCR, and AC2 results were confirmed by using different NAATs on the original specimen. For FCU samples from men, any NAAT can be used for confirmation. However, for all other specimen types, some NAATs cannot be used to confirm positive results from other NAATs. Thus, a single repeat test appears to be a reliable method for confirmation, but by doing more extensive testing, an additional 5% were confirmed. With >90% of all GC-positive NAATs being confirmed, our results show that confirmatory testing is not warranted for these genital specimens.