Abstract
To study the molecular characteristics of Chlamydia trachomatis, the major outer membrane protein gene (ompA) of C. trachomatis from primary school students with trachoma residing in the Qinghai Tibetan area was sequenced and compared with the same serotype in GenBank. In Jianshetang Primary School and Galeng Central Primary School in the Galeng Tibetan Township of Qinghai Haidong Sala Autonomous County, scraped samples were collected from the upper tarsal conjunctiva and lower conjunctival sac of both eyes of 45 students with trachoma, stored at 4°C, and transported to Beijing Tongren Hospital by air within 24 h. The samples were screened for C. trachomatis by real-time PCR. The ompA gene from the C. trachomatis-positive samples was amplified by nested PCR. The serotype was confirmed by National Center for Biotechnology Information (NCBI) BLAST search and homology analysis. The entire ompA gene sequence was compared with the corresponding gene sequences of serotype B strains available in GenBank. Of the 45 students aged 6–13 years with trachoma, 26 C. trachomatis-positive students were identified by the initial real-time PCR screening (average age, (9.09±1.63) years; sex ratio, 1.0), accounting for 57.78% (26/45). The cycle threshold values for real-time PCR were 16.79–37.77. Half (13/26) of C. trachomatis-positive students had a bacterial copy number of >105. The compliance rate of the ompA gene sequences with the C. trachomatis serotype B strains in GenBank was up to 99%. Two novel genetic mutations were found when the ompA gene was compared with those of the 11 serotype B strains in GenBank. The two non-synonymous mutations were located at (i) position 271 in the second constant domain, an adenine (A) to guanine (G) substitution (ACT→GCT), changing the amino acid at position 91 from threonine to alanine (Thr→Ala) in all 26 strains; and (ii) position 887 in the fourth variable domain, a cytosine (C) to thymine (T) substitution (GCA→GTA), changing the amino acid at residue 296 from alanine to valine (Ala→Val) in four of the 26 strains. Six mutations were identified relative to ATCC VR-573. The strains could be divided into two gene clusters according to the mutation at nucleotide position 887: CQZ-1 (China Qinghai Tibetan-1) and CQZ-2 (China Qinghai Tibetan-2). We thus detected two novel serotype B mutant strains of C. trachomatis among study subjects with trachoma.
![](http://media.springernature.com/lw685/springer-static/image/art%3A10.1007%2Fs11427-016-5059-9/MediaObjects/11427_2016_5059_Fig1_HTML.jpg)
![](http://media.springernature.com/lw685/springer-static/image/art%3A10.1007%2Fs11427-016-5059-9/MediaObjects/11427_2016_5059_Fig2_HTML.jpg)
![](http://media.springernature.com/lw685/springer-static/image/art%3A10.1007%2Fs11427-016-5059-9/MediaObjects/11427_2016_5059_Fig3_HTML.jpg)
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Baehr, W., Zhang, Y., Joseph, T., Su, H., Nano, F.E., Everett, K.D., and Caldwell, H.D. (1988). Mapping antigenic domains expressed by Chlamydia trachomatis major outer membrane protein genes. Proc Natl Acad Sci USA 85, 4000–4004.
Bandea, C.I, Kubota, K., Brown, T.M., Kilmarx, P.H., Bhullar, V., Yanpaisarn, S., Chaisilwattana, P., Siriwasin, W., and Black, C.M. (2001). Typing of Chlamydia trachomatis strains from urine samples by amplification and sequencing the major outer membrane protein gene (omp1). Sex Transm Infect 77, 419–422.
Batteiger, B.E., Tu, W., Ofner, S., Van Der Pol, B., Stothard, D.R., Orr, D.P., Katz, B.P., and Fortenberry, J.D. (2010). Repeated Chlamydia trachomatis genital infections in adolescent women. J Infect Dis 201, 42–51.
Seth-Smith, H.M., Harris, S.R., Persson, K., Marsh, P., Barron, A., Bignell, A., Bjartling, C., Clark, L., Cutcliffe, L.T., Lambden, P.R., Lennard, N., Lockey, S.J., Quail, M.A., Salim, O., Skilton, R.J., Wang, Y., Holland, M.J., Parkhill, J., Thomson, N.R., and Clarke, I.N. (2009). Co-evolution of genomes and plasmids within Chlamydia trachomatis and the emergence in Sweden of a new variant strain. BMC Genomics 10, 239.
Brunelle, B.W., and Sensabaugh, G.F. (2006). The ompA gene in Chlamydia trachomatis differs in phylogeny and rate of evolution from other regions of the genome. Infect Immun 74, 578–585.
Burton, M.J., Holland, M.J., Makalo, P., Aryee, E.A., Sillah, A., Cohuet, S., Natividad, A., Alexander, N.D., Mabey, D.C., and Bailey, R.L. (2010). Profound and sustained reduction in Chlamydia trachomatis in The Gambia: a five-year longitudinal study of trachoma endemic communities. PLoS Negl Trop Dis 4, 10.
Harris, S.R., Clarke, I.N., Seth-Smith, H.M., Solomon, A.W., Cutcliffe, L.T., Marsh, P., Skilton, R.J., Holland, M.J., Mabey, D., Peeling, R.W., Lewis, D.A., Spratt, B.G., Unemo, M., Persson, K., Bjartling, C., Brunham, R., de Vries, H.J., Morré, S.A., Speksnijder, A., Bébéar, C.M., Clerc, M., de Barbeyrac, B., Parkhill, J., and Thomson, N.R. (2012). Whole-genome analysis of diverse Chlamydia trachomatis strains identifies phylogenetic relationships masked by current clinical typing. Nat Genet 44, 413–419, S411.
Hayes, L.J, Pickett, M.A., Conlan, J.W., Ferris, S., Everson, J.S., Ward, M.E., and Clarke, I.N. (1990). The major outer-membrane proteins of Chlamydia trachomatis serovars A and B: intra-serovar amino acid changes do not alter specificities of serovar- and C subspecies-reactive antibody-binding domains. J Gen Microbiol 136, 1559–1566.
Hsieh, Y.H., Bobo, L.D., Quinn, T.C., and West, S.K. (2001). Determinants of trachoma endemicity using Chlamydia trachomatis ompA DNA sequencing. Microbes Infect 3, 447–458.
Ishak, Mde.O., Costa, M.M., de Almeida, N.C., Santiago, A.M., de Brito, W.B., Vallinoto, A.C., Azevedo, V.N., and Ishak, R. (2015). Chlamydia trachomatis serotype A infections in the Amazon region of Brazil: prevalence, entry and dissemination. Rev Soc Bras Med Trop 48, 170–174.
Joseph, S.J., Didelot, X., Rothschild, J., de Vries, H.J., Morre, S.A., Read, D., and Dean, D. (2012). Population genomics of Chlamydia trachomatis: insights on drift, selection, recombination, and population structure. Mol Biol Evol 29, 3933–3946.
King, J.D., Odermatt, P., Utzinger, J., Ngondi, J., Bamani, S., Kamissoko, Y., Boubicar, K., Hassan, A.S., Nwobi, B.C., Jip, N., Amnie, A., Teferi, T., Mosher, A.W., Stewart, A.E., Cromwell, E.A., and Emerson P.M. (2013). Trachoma among children in community surveys from four African countries implications of using school surveys for evaluating prevalence. Int Health 5, 280–287.
Last, A.R., Burr, S.E., Weiss, H.A., Harding-Esch, E.M., Cassama, E., Nabicassa, M., Mabey, D.C., Holland, M.J., and Bailey, R.L. (2014). Risk factors for active trachoma and ocular Chlamydia trachomatis infection in treatment-naive trachoma-hyperendemic communities of the Bijagos Archipelago, Guinea Bissau. PLoS Negl Trop Dis 8, 2900.
Mital, J., Miller, N.J., Dorward, D.W., Dooley, C.A., and Hackstadt, T. (2013). Role for chlamydial inclusion membrane proteins in inclusion membrane structure and biogenesis. PLoS One 8, 63426.
Naito, T., Aoki, K., Ohguchi, T., Ohgami, K., Ohno, S., Shiot, H., Shrestha, J.K., and Upadhyay, M.P. (2009). Pathogenesis of infectious conjunctivitis in Nepal. Nippon Ganka Gakkai Zasshi 113, 1088–1091.
Nigusie, A., Berhe, R., and Gedefaw, M. (2015). Prevalence and associated factors of active trachoma among children aged 1–9 years in rural communities of GonjiKolella district, West Gojjam zone, North West Ethiopia. BMC Res Notes 8, 641.
Nunes, A., Borrego, M.J., Nunes, B., Florindo, C., and Gomes, J.P. (2009). Evolutionary dynamics of ompA, the gene encoding the Chlamydia trachomatis key antigen. J Bacteriol 191, 7182–7192.
Porter, M., Mak, D., Chidlow, G., Harnett, G.B., and Smith, D.W. (2008). The molecular epidemiology of ocular Chlamydia trachomatis infections in Western Australia: implications for trachoma control. Am J Trop Med Hyg 78, 514–517.
Seth-Smith, H.M., Harris, S.R., Persson, K., Marsh, P., Barron, A., Bignell, A., Hayes, L.J., Pickett, M.A., Conlan, J.W., Ferris, S., Everson, J.S., Ward, M.E., and Clarke, I.N. (1990). The major outer-membrane proteins of Chlamydia trachomatisserovars A and B: intra-serovar amino acid changes do not alter specificities of serovar- and C subspecies-reactive antibody-binding domains. J Gen Microbiol 136, 1559–1566.
Shattock, A.J., Gambhir, M., Taylor, H.R., Cowling, C.S., Kaldor, J.M., and Wilson, D.P. (2015). Control of trachoma in Australia: a model based evaluation of current interventions. PLoS Negl Trop Dis 9, e0003474.
Stephens, R.S., Sanchez-Pescador, R., Wagar, E.A., Inouye, C., and Urdea, M.S. (1987). Diversity of Chlamydia trachomatis major outer membrane protein genes. J Bacteriol 169, 3879–3885.
Takahashi, S., Yamazaki, T., Satoh, K., Inoue, M., Takahashi, S., Ishihara, O., Oka, Y., Horiguchi, Y., Okuwaki, Y., Suzuki, S., and Kishimoto, T. (2007). Longitudinal epidemiology of Chlamydia trachomatisserovars in female patients in Japan. Jpn J Infect Dis 60, 374–376.
Takourt, B., de Barbeyrac, B., Khyatti, M., Radouani, F., Bebear, C., Dessus-Babus, S., Bebear, C., and Benslimane, A. (2001). Direct genotyping and nucleotide sequence analysis of VS1 and VS2 of the Omp1 gene of Chlamydia trachomatis from Moroccan trachomatous specimens. Microbes Infect 3, 459–466.
Taylor, H.R., Burton, M.J., Haddad, D., West, S., and Wright, H. (2014). Trachoma. Lancet 384, 2142–2152.
Thylefors, B., Dawson, C.R., Jones, B.R., West, S.K., and Taylor, H.R. (1987). A simple system for the assessment of trachoma and its complications. Bull World Health Organ 65, 477–483.
Wang, N., Hu, A., and Taylor, H.R. (2015). Trachoma. (Beijing: People’s Medical Publishing House), pp. 50.
Author information
Authors and Affiliations
Corresponding author
Additional information
This article is published with open access at link.springer.com
Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
An erratum to this article can be found at http://dx.doi.org/10.1007/s11427-016-5097-3.
Rights and permissions
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0), which permits use, duplication, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Li, X., Zhang, S., Liang, Q. et al. Molecular characteristics of the ompA gene of serotype B Chlamydia trachomatis in Qinghai Tibetan primary school students. Sci. China Life Sci. 59, 561–570 (2016). https://doi.org/10.1007/s11427-016-5059-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-016-5059-9