Clinical and analytical evaluation of the new Aptima Mycoplasma genitalium assay, with data on M. genitalium prevalence and antimicrobial resistance in M. genitalium in Denmark, Norway and Sweden in 2016

Unemo, Mats; Salado-Rasmussen, Kirsten; Hansen, Marit; Olsen, Anne O.; Falk, My; Golparian, Daniel; Aasterød, Marie; Ringlander, Johan; Nilsson, Christian; Sundqvist, Martin; Schønning, Kristian; Moi, Harald; Westh, Henrik; Jensen, J S

doi:10.1016/j.cmi.2017.09.006

Cited by 85 publications

(96 citation statements)

References 33 publications

(32 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…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: 98%

“…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%

See 1 more Smart Citation

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

et al. 2019

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

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

et al. 2019

View full text Add to dashboard Cite

show abstract

“…This might explain the difference in sensitivity of the MgPa assay and Mg Macrolide R assay. Using RNA as input obviously increases the sensitivity when compared to DNA as used in our RTN-qPCR sequencing method and is employed by Aptima PCRs [30,31].…”

Section: Discussionmentioning

confidence: 99%

Sensitive and specific assay for the simultaneous detection of Mycoplasma genitalium and macrolide resistance-associated mutations

Braam

Marm

Severs

et al. 2018

Eur J Clin Microbiol Infect Dis

View full text Add to dashboard Cite

Patients infected by Mycoplasma genitalium are often treated empirically with the macrolide azithromycin. Macrolide resistance is becoming quite common; empirical treatment is compromised. Sequencing was initially used to detected azithromycin resistance-associated mutations. As this was laborious, qPCRs have been developed for their detection. In the present study, we describe a fast, sensitive, and specific qPCR assay that enables routine testing of M. genitalium and macrolide resistanceassociated mutations in a single assay. M. genitalium positive clinical samples were used to compare (i) the commonly used MgPa assay for the detection of M. genitalium infections (MgPa qPCR), (ii) a combined 23S rRNA gene PCR/sequencing assay (Mg23S qPCR/Sequencing) to identify macrolide resistance-associated mutations, and (iii) our newly developed probe-based melt curve qPCR for simultaneous detection of M. genitalium and macrolide resistance-associated mutations (Macrolide-R/MG ELITe MGB Kit, Elitech Bothel USA in short Mg Macrolide R qPCR). Specificity of the qPCR was tested using urogenital samples that were tested positive for a range of other microorganisms. M. genitalium was detected in 196/236 (83.1%) samples by the MgPa qPCR, versus 172/236 (72.9%) by the combined Mg23S qPCR/Sequencing, and 202/236 (85.6%) by the Mg Macrolide R qPCR. The Mg Macrolide R qPCR showed high concordance to the Mg23S qPCR/Sequencing assay (201 vs 202 could be genotyped, respectively) for the detection of the macrolide resistant mutations. None of the other urogenital pathogens were tested positive in the Mg Macrolide R qPCR, indicating specificity. The Mg Macrolide R qPCR is fast, sensitive, specific, and can easily be implemented in the routine diagnostics.

show abstract

“…Распространенность резистентных мутаций для азитромицина и моксифлоксацина составляла 41,4 (17,7-56,6) и 6,6% (4,1-10,2%) соответственно. Также в этих странах была обнаружена множественная лекарственная устойчивость (2,7; 1,1-4,2%) [59].…”

Section: проблема антибиотикорезистентности M Genitaliumunclassified

Current state of the research on Mycoplasma genitalium infection

Потекаев¹,

Kisina²,

Romanova³

et al. 2018

Klin. dermatol. venerol.

View full text Add to dashboard Cite

Характерные особенности Mycoplasma genitaliumМикоплазмы являются наименьшими по размерам прокариотами, способными самостоятельно размножаться. Они принадлежат к классу Mollicutes («мягкая кожа») и эволюционировали регрессивно путем сокращения генома грамположительных бактерий предшественников [1]. Генетическую информацию обеспечивает геном, который в случае Mycoplasma genitalium является наименьшим известным из самореплицирующихся структур [2]. В течение многих лет молликуты относили к вирусам, так как они способны проходить через фильтры с диаметром пор 0,45 и даже 0,22 мкм. В 30-х годах XX века, когда концепция вирусов была более четко сформулирована, микоплазмы стали относить к бактериям. В 50-х годах некоторые виды микоплазм относили к L-формам бактерий -бактерий без клеточной стенки, и только в 60-х годах микоплазмы заняли то место в таксономии, которое они занимают и сейчас. Проходимость через бактериологические 1 ГБУЗ Москвы «Московский научно-практический центр дерматовенерологии и косметологии» Департамента здравоохранения Москвы, Москва, Россия; 2 ФБУН «Центральный научно-исследовательский институт эпидемиологии Роспотребнадзора», Москва, Россия В настоящее время Mycoplasma genitalium признана безусловным патогеном, в связи с чем обнаружение данного микроорганизма служит абсолютным показанием для назначения этиотропной терапии. Лечение больных с инфекцией M. genitalium предотвращает дальнейшую передачу возбудителя и развитие осложнений, включающих воспалительные заболевания органов малого таза и трубное бесплодие. Одной из современных ключевых проблем лечения инфекции M. genitalium является стремительное развитие резистентности этого возбудителя к антибактериальным препаратам различных фармакологических групп по всему миру.Ключевые слова: Mycoplasma genitalium, обзор, клиника, лечение, резистентность. Currently, Mycoplasma genitalium is recognized as a non-opportunistic pathogen, and therefore detection of this pathogen is an absolute indication for causal treatment. Treatment of patients with M. genitalium infection prevent further transmission of the causative agent and the development of complications, including pelvic inflammatory disease and tubal infertility. The rapid development of the resistance of the pathogen to antibacterial drugs of various pharmacological groups around the world is one of the key current challenges in treatment of M. genitalium infection.

show abstract

Clinical and analytical evaluation of the new Aptima Mycoplasma genitalium assay, with data on M. genitalium prevalence and antimicrobial resistance in M. genitalium in Denmark, Norway and Sweden in 2016

Cited by 85 publications

References 33 publications

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

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

Sensitive and specific assay for the simultaneous detection of Mycoplasma genitalium and macrolide resistance-associated mutations

Current state of the research on Mycoplasma genitalium infection

Contact Info

Product

Resources

About