Application of an oligonucleotide array assay for rapid detecting and genotyping of Chlamydia trachomatis from urogenital specimens

Zheng, Heping; Jiang, Lifang; Fang, D.Z.; Xue, Yaohua; Wu, Ya-an; Huang, Jinmei; Ou, Zhanpeng

doi:10.1016/j.diagmicrobio.2006.05.007

Cited by 26 publications

(40 citation statements)

References 20 publications

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“…Importantly, these mixed preparations were arbitrarily prepared with a 100-fold difference in bacterial load, and it should be emphasized that a natural mixed infection could comprise a severalfold-higher difference between coinfecting serovars, which could ultimately affect the detection of a serovar at lower levels. The frequency of multiple serovars identified in this study (2.4%) is comparable to those in other C. trachomatis typing studies of between 1.5% and 5.4% (8,15,19,20,33). Interestingly, some studies have described multiple infections with frequencies as high as 8.7% and 13% (14,31) which principally involved use of PCR followed by reverse line blot procedures.…”

Section: Discussionsupporting

confidence: 68%

“…With this, a notable limitation of this qPCR assay was its marginally lower success rate in predicting C. trachomatis serovars compared to that of sequencing. Several of the published protocols for C. trachomatis serovar prediction incorporate an initial nested PCR followed by various methods of detection, including sequencing, qPCR, blotting, and microsphere suspension array, to improve assay sensitivity (7,8,9,14,31,33). Therefore, as a step to potentially enhancing the current assay, it is possible that a nested PCR precedes the primary consensus/group-specific PCR.…”

Section: Discussionmentioning

confidence: 99%

“…C. trachomatis serovars can be subdivided into three distinct phylogenetic clades based on the ompA gene: the B group (comprising B/Ba, D, E, L1, and L2), the C group (comprising A, C, H, I, J, K, and L3), and the intermediate (I) group (comprising F and G). Although there is no correlation of phylogenetic clades with characteristic disease, these clades/groups have value for the development of more rapid screening assays for predicting serovars (19,20,33).…”

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confidence: 99%

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Development and Evaluation of anompAQuantitative Real-Time PCR Assay forChlamydia trachomatisSerovar Determination

et al. 2010

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Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Development and Evaluation of anompAQuantitative Real-Time PCR Assay forChlamydia trachomatisSerovar Determination

et al. 2010

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“…All primers and probes were synthesized by Metabion International AGi, Germany, except the MGB-labeled probes, which were synthesized by Applied Biosystems, Cheshire, United Kingdom ( Table 1). The two-step nested PCR, as used in this study and in previously published studies (1,7,8,9,10,15,20,21,22), carries an inherent risk of contaminating equipment and laboratory environment. Stringent physical separation of different steps in the assays, namely, the use of dedicated equipment for each step, a one-way flow of work from extraction to amplification, meticulous techniques to prevent the production of aerosols from tubes containing amplicons, and filtered tips avoided this problem.…”

Section: Methodsmentioning

confidence: 99%

“…The amplicons are being used for typing by restriction fragment length polymorphism (RFLP) analysis (2,7,9,12), DNA sequencing (1,8,15,21), reverse line blot hybridization (10,16,20), and oligonucleotide arrays (22). These studies have resulted in more than 60 reports since 1991 according to PubMed.…”

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confidence: 99%

Development of Real-Time PCR Assays for Genotyping of Chlamydia trachomatis

Jalal

Stephen

Alexander³

et al. 2007

J Clin Microbiol

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We have developed and validated a nested real-time PCR (NRT-PCR) for the genotyping of Chlamydia trachomatis and used it specifically for the typing of either eight genovars from D to K or three genovars of lymphogranuloma venereum (LGV). The 11 probes used in the NRT-PCR correctly identified the DNA from D to K and LGV reference strains and did not cross-react with the DNA from 26 strains representing the bacterial pathogens and commensals of the oropharynx, genital tract, and rectum. The NRT-PCR had a 95% probability of detection at four genome copies (confidence interval, three to six copies) of C. trachomatis per reaction. One hundred cervical and urethral swab specimens containing C. trachomatis DNA from 63 women and 37 men were used to validate the method. The results from the NRT-PCR and the DNA sequencing of amplicons generated from the omp1 gene showed 100% correlation for these samples. The assay also identified the LGV-II genotype in 24 of 48 rectal swab specimens containing C. trachomatis DNA that were obtained from men having sex with men. The Sexually Transmitted Bacteria Reference Laboratory, London, independently confirmed these results using group-specific LGV real-time PCR and restriction fragment length polymorphism analysis. Compared with the NRT-PCR, non-NRT-PCR was found to be less sensitive: it typed C. trachomatis DNA in only 80% of the genital samples and 90% of the rectal swab samples. This is the first successful demonstration of the use of real-time PCR for the genotype-specific typing of C. trachomatis strains that cause sexually transmitted diseases.

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Comparison of reverse hybridization and ompA sequencing methods applied on Chlamydia trachomatis strains from Tunisia

et al. 2017

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Two techniques based on ompA amplification of Chlamydia trachomatis were compared, being reverse hybridization (RHM) and ompA sequencing (OSA), to investigate the concordance between them and to study the epidemiological relevance of each method. In addition, phylogenetic analysis was performed on the ompA sequences. One hundred and seven C. trachomatis positive samples from Tunisian patients and female sex workers were analyzed using both the RHM and ompA sequencing. The overall genovar distribution obtained with both techniques was very similar. The RHM identified nine genovars, being B, D, E, F, G, H, I, J and K, where B, I, J, and K were only found in mixed infections versus 7 types for the OSA being D, E, F, G, H, I, and K. The agreement between both typing techniques was 87.8%. Both methods showed that genovar E was the most predominant type. In 24.3% of the analyzed samples, mixed infections were detected. In 96.1% of these, the genovar identified by OSA was also detected using the RHM. OmpA sequencing allowed determination of six genovar types that could not be typed using RHM. The analyses of ompA nucleotide variation in the 107 clinical specimens detected ompA genovar variants with distinct ompA mutational patterns for types D2, G1, G2, and H1. In conclusion, RHM and OSA showed a high agreement in C. trachomatis genotyping results with each having their specific benefits.

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Application of an oligonucleotide array assay for rapid detecting and genotyping of Chlamydia trachomatis from urogenital specimens

Cited by 26 publications

References 20 publications

Development and Evaluation of anompAQuantitative Real-Time PCR Assay forChlamydia trachomatisSerovar Determination

Development and Evaluation of anompAQuantitative Real-Time PCR Assay forChlamydia trachomatisSerovar Determination

Development of Real-Time PCR Assays for Genotyping of Chlamydia trachomatis

Comparison of reverse hybridization and ompA sequencing methods applied on Chlamydia trachomatis strains from Tunisia

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