Evaluation of the Hologic Panther Transcription-Mediated Amplification Assay for Detection of Mycoplasma genitalium

Tabrizi, Sepehr N.; Costa, Anna‐Maria; Su, Jenny; Lowe, P; Bradshaw, Catriona; Fairley, CK; Garland, Suzanne M.

doi:10.1128/jcm.01038-16

Cited by 25 publications

(21 citation statements)

References 21 publications

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“…Previous studies in Sydney and Melbourne, of 508 and 515 men, respectively, both failed to detect pharyngeal Mgen using PCR. TMA has a higher analytical sensitivity than PCR for Mgen,27 although the Aptima Mycoplasma genitalium Assay (Hologic) has yet to be validated for detection of pharyngeal Mgen. Given the organism load of Mgen is up to 100 times lower than that of CT,28 and may be at particularly low loads in the pharynx, TMA may be more likely to detect pharyngeal infections.…”

Section: Discussionmentioning

confidence: 99%

ExtragenitalMycoplasma genitaliuminfections among men who have sex with men

et al. 2019

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ObjectivesThere are limited data on the prevalence of Mycoplasma genitalium (Mgen) coinfection with rectal chlamydia (Chlamydia trachomatis (CT)) and rectal gonorrhoea (Neisseria gonorrhoeae (NG)) infections and few studies examining the prevalence of pharyngeal Mgen in men who have sex with men (MSM). Using transcription-mediated amplification assay, this study aimed to determine the proportion of rectal CT and rectal NG infections in MSM who are coinfected with rectal Mgen, and the proportion of MSM with Mgen detected in the pharynx in order to inform clinical practice.MethodsThis was a cross-sectional study conducted at Melbourne Sexual Health Centre in Australia. Consecutively collected rectal swabs from MSM that tested positive for CT (n=212) or NG (n=212), and consecutively collected pharyngeal samples (n=480) from MSM were tested for Mgen using the Aptima Mycoplasma genitalium Assay (Hologic, San Diego). Samples were linked to demographic data and symptom status.ResultsRectal Mgen was codetected in 27 of 212 rectal CT (13%, 95% CI 9 to 18) and in 29 of 212 rectal NG (14%, 95% CI 9 to 19) samples, with no difference in the proportion positive for Mgen. MSM with rectal CT/Mgen coinfection had more sexual partners than those with rectal CT monoinfection (mean 6 vs 11, p=0.06). MSM with rectal NG/Mgen coinfection were more likely to be HIV-positive than those with rectal NG monoinfection (OR=2.96, 95% CI 1.21 to 7.26, p=0.023). MSM with rectal CT/Mgen coinfection were more likely to be using pre-exposure prophylaxis than MSM with rectal NG/Mgen coinfection (OR 0.25, 95% CI 0.10 to 0.65, p=0.002). Pharyngeal Mgen was uncommon and detected in 8 of 464 samples (2%, 95% CI 1% to 3%). Pharyngeal Mgen was associated with having a rectal STI (OR=10.61, 95% CI 2.30 to 48.87, p=0.002), and there was a borderline association with being HIV-positive (p=0.079).ConclusionThese data indicate one in seven MSM treated for rectal CT or rectal NG will have undiagnosed Mgen that is potentially exposed to azithromycin during treatment of these STIs. Rectal Mgen coinfection was associated with specific risk factors which may inform testing practices. Pharyngeal Mgen was uncommon.

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Section: Discussionmentioning

confidence: 99%

ExtragenitalMycoplasma genitaliuminfections among men who have sex with men

et al. 2019

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“…Several companies have commercialized CE-marked multiplex PCR assays for the detection of sexually transmitted pathogens, including M. genitalium ( 6 , 7 , 9 , 10 ). As an alternative to PCR, Hologic, Inc. developed a research-use-only (RUO) TMA assay for the detection of M. genitalium ( 8 , 11 , 12 ) and recently commercialized the Aptima Mycoplasma genitalium TMA (MG-TMA) assay CE-marked for in vitro diagnosis (CE-IVD), which is available on the Panther platform. Commercial tests that have been CE-marked for document conformity usually suffer from limited clinical validation.…”

Section: Introductionmentioning

confidence: 99%

French Prospective Clinical Evaluation of the Aptima Mycoplasma genitalium CE-IVD Assay and Macrolide Resistance Detection Using Three Distinct Assays

et al. 2017

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The aim of this study was to evaluate the clinical performance of the Aptima Mycoplasma genitalium transcription-mediated amplification (MG-TMA) CE-marked for in vitro diagnosis (CE-IVD) assay for the detection of Mycoplasma genitalium in male and female clinical samples in comparison with the in-house real-time PCR (in-house PCR) assay routinely used in our laboratory. A total of 1,431 clinical specimens obtained from 1,235 patients were prospectively collected at the Bacteriology Department of Bordeaux University Hospital (France). Additional research-use-only Aptima M. genitalium transcription-mediated amplification (TMA) assays, Alt1-TMA and Alt2-TMA, were performed on discordant specimens to determine M. genitalium infection status. All confirmed M. genitalium-positive specimens were tested for macrolide resistance using three assays: the in-house 23S rRNA FRET PCR assay, the SpeeDx ResistancePlus MG assay and the nested reverse transcription-PCR (RT-PCR) sequencing assay. The comparison of the MG-TMA assay with the in-house PCR results showed a moderate correlation (kappa value, 0.69). The MG-TMA assay had higher clinical sensitivity compared to that of the in-house PCR assay (100% versus 59.74%, respectively) and similar specificity (99.10% versus 100%, respectively) for M. genitalium detection. In this study, the prevalence of M. genitalium infection was 5.90% (72/1,220 patients). The nested RT-PCR sequencing assay was the most sensitive but the most laborious assay for detecting macrolide-resistance-associated mutations. The prevalence of resistance was 8.33% (6/72). To our knowledge, this is the first clinical evaluation of the MG-TMA CE-IVD assay. The MG-TMA assay performed on the automated Panther system is a very sensitive and specific method for the detection of M. genitalium in clinical specimens.

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“…In this study, we validated the performance characteristics of four TMA NAAT assays for the detection of ribosomal RNAs from M. genitalium. We found that these assays are sensitive and specific for the detection of multiple stains of M. genitalium and have Similarly to NAATs designed to detect other sexually transmitted infections such as Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis, the detection of rRNA of M. genitalium affords a particularly sensitive method for detecting the organism in vivo, leading to improved clinical diagnosis of M. genitalium infection compared to that with NAATs that target genomic DNA of the organism (20)(21)(22). This difference in relative test clinical sensitivity may be due to the disparity between genomic DNA and rRNA content in the cell (i.e., a single DNA genome versus an estimated hundreds to thousands of rRNA copies per cell) (29), enabling the detection of low titers of M. genitalium in patient specimens (30,31).…”

Section: Discussionmentioning

confidence: 97%

“…This is the first study to use only rRNA-specific NAATs for the components of a composite comparator reference standard for the evaluation of an rRNA-based molecular test for M. genitalium detection. Previous studies investigating the clinical perfor- mance of rRNA-based NAAT assays for M. genitalium used a reference standard consisting of a combination of tests that detected rRNA and genomic DNA of M. genitalium (20,21). This mixed reference standard approach has the potential to introduce bias into the analyses, since the lower sensitivity of DNA-based tests can decrease the rate of DNA-positive/RNA-positive consensus results obtained in the analysis, potentially resulting in artificially lowered specificity estimates for the RNA-based diagnostic test under evaluation.…”

Section: Discussionmentioning

confidence: 99%

“…The TMA-based Aptima Mycoplasma genitalium assay (AMG) is an FDA-cleared (510k number DEN180047) in vitro diagnostic (IVD) test for the detection of 16S rRNA from M. genitalium. Application of the AMG IVD assay outside the United States has been described (20)(21)(22). In the present study, we developed and optimized three additional alternate (Alt) TMA assays, Alt TMA-1, Alt TMA-2, and Alt TMA-3, to capture, amplify, and detect unique regions of 16S rRNA (Alt TMA-1) or 23S rRNA (Alt TMA-2 and Alt TMA-3) of M. genitalium.…”

Section: Methodsmentioning

confidence: 99%

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Design and Validation of Transcription-Mediated-Amplification Nucleic Acid Amplification Tests for Mycoplasma genitalium

et al. 2019

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Mycoplasma genitalium is a sexually transmitted bacterium linked to adverse sexual and reproductive health outcomes in women and men. M. genitalium is difficult to culture, and in the absence of validated amplified molecular methods for diagnosis of infection, there is no reference standard available for use as a comparator for the validation of new M. genitalium diagnostic tests. We evaluated the analytical and clinical performance of three transcription-mediated amplification (TMA) tests for M. genitalium, each targeting unique rRNA sequences, for use as a composite comparator for clinical validation of the Aptima Mycoplasma genitalium (AMG) assay, an in vitro diagnostic (IVD) TMA test that targets 16 s rRNA of M. genitalium. Analytical sensitivity, specificity, and strain inclusivity of all four TMA tests were determined using nine laboratory strains of M. genitalium and 56 nontarget bacteria, protozoa, and viruses. Analytical sensitivity of the tests for M. genitalium ranged from 0.017 to 0.040 genome equivalents/ml. None of the nontarget organisms evaluated cross-reacted with any test. A composite comparator reference standard consisting of the 3 alternate (Alt) TMA tests was used to evaluate the clinical performance of the AMG assay by testing residual vaginal swab, female urine, and male urine specimens obtained from 1,400 adult subjects from three U.S. clinical sites. Using this reference standard to establish infected specimen status, the sensitivity, specificity, and overall agreement of the AMG IVD assay were 100%, 99.9%, and 99.9%, respectively. These results demonstrate the utility of molecular composite reference standard methodology for the clinical validation of future IVD tests for this organism.

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Evaluation of the Hologic Panther Transcription-Mediated Amplification Assay for Detection of Mycoplasma genitalium

Cited by 25 publications

References 21 publications

ExtragenitalMycoplasma genitaliuminfections among men who have sex with men

ExtragenitalMycoplasma genitaliuminfections among men who have sex with men

French Prospective Clinical Evaluation of the Aptima Mycoplasma genitalium CE-IVD Assay and Macrolide Resistance Detection Using Three Distinct Assays

Design and Validation of Transcription-Mediated-Amplification Nucleic Acid Amplification Tests for Mycoplasma genitalium

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