Abstract
The identification of Chlamydia trachomatis genotypes is important for both molecular epidemiology and infection control such as contact tracing and identification of high-risk groups. Currently, at least 19 human serovars have been recognized by using polyclonal and monoclonal antibodies against the major outer membrane protein. In sexually transmitted diseases, multiple pathogens or genotype infections are not uncommon. Hence, detection of multiple gene targets in one reaction is becoming increasingly important. Here, we describe the multiplex detection of eight genotypes of C. trachomatis by a combination of a PCR amplification with a multiplex bead array detection. The bead array system comprises distinct bead sets, which are color coded by different fluorescent intensities and a dual-laser flow cytometer analyzer to identify the identity of the bead and the intensity of the reporter dye that binds to the target molecules. The DNA sequences of the variable segments (VS2 or VS1–VS2) in outer membrane protein (omp1) gene are PCR amplified and biotin labeled and used as a gene target for the genotyping of C. trachomatis. Genotype-specific probes coupled to beads are used for capturing the labeled target amplicons through specific hybridization. Thus, multiple genotypes are detected and differentiated simultaneously by yielding quantitative data.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gaydos CA, Theodore M, Dalesio N, Wood BJ, Quinn TC (2004) Comparison of three nucleic acid amplification tests for detection of Chlamydia trachomatis in urine specimens. J Clin Microbiol 42:3041–3045
Eckert LO, Suchland RJ, Hawes SE, Stamm WE (2000) Quantitative Chlamydia trachomatis cultures: correlation of chlamydial inclusion-forming units with serovar, age, sex, and race. J Infect Dis 182:540–544
Suchland RJ, Stamm WE (1991) Simplified microtiter cell culture method for rapid immunotyping of Chlamydia trachomatis. J Clin Microbiol 29:1333–1338
Yuan Y, Zhang YX, Watkins NG, Caldwell HD (1989) Nucleotide and deduced amino acid sequences for the four variable domains of the major outer membrane proteins of the 15 Chlamydia trachomatis serovars. Infect Immun 57:1040–1049
Wang SP, Kuo CC, Barnes RC, Stephens RS, Grayston JT (1985) Immunotyping of Chlamydia trachomatis with monoclonal antibodies. J Infect Dis 152:791–800
Zheng HP, Jiang LF, Fang DY, Xue YH, Wu YA, Huang JM, Ou ZY (2007) Application of an oligonucleotide array assay for rapid detecting and genotyping of Chlamydia trachomatis from urogenital specimens. Diagn Microbiol Infect Dis 57:1–6
Jalal H, Stephen H, Alexander S, Carne C, Sonnex C (2007) Development of real-time PCR assays for genotyping of Chlamydia trachomatis. J Clin Microbiol 45:2649–2653
Xiong L, Kong F, Zhou H, Gilbert GL (2006) Use of PCR and reverse line blot hybridization assay for rapid simultaneous detection and serovar identification of Chlamydia trachomatis. J Clin Microbiol 44:1413–1418
Molano M, Meijer CJ, Morre SA, Pol R, van den Brule AJ (2004) Combination of PCR targeting the VD2 of omp1 and reverse line blot analysis for typing of urogenital Chlamydia trachomatis serovars in cervical scrape specimens. J Clin Microbiol 42:2935–2939
Anderson M, Handley J, Hopwood L, Murant S, Stower M, Maitland NJ (1997) Analysis of prostate tissue DNA for the presence of human papillomavirus by polymerase chain reaction, cloning, and automated sequencing. J Med Virol 52:8–13
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Huang, CT., Li, SY. (2012). Protocol for the Use of a Bead Array for the Multiple Detection of Genotype of Chlamydia trachomatis . In: MacKenzie, C., Henrich, B. (eds) Diagnosis of Sexually Transmitted Diseases. Methods in Molecular Biology, vol 903. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-937-2_12
Download citation
DOI: https://doi.org/10.1007/978-1-61779-937-2_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-936-5
Online ISBN: 978-1-61779-937-2
eBook Packages: Springer Protocols