Gene typing of chlamydia trachomatis by polymerase chain reaction and restriction endonuclease digestion

Charlotte A Gaydos, Linda Bobo, Laura Welsh, Edward W. Hook, Raphael P Viscidi, Thomas C Quinn

Research output: Contribution to journalArticle

Abstract

A portion of the major outer membrane protein (MOMP) gene from 15 Chlamydia trachomatis serovars was amplified by polymerase chain reaction (PCR) and the product was analyzed by restriction fragment length polymorphism (RFLP). A set of primers was used to amplify an 871 base pair gene fragment encompassing the 4 hypervariable regions of MOMP. AluI digestion of the product gave distinctive patterns for the 15 serovars as demonstrated on silver-stained polyacrylamide gels. A triple digest with EcoRI, HinfI, and HpaII allowed improved discrimination between closely related serovars (C, H, I, J, L3). PCR and RFLP were used to type 50 wild-type clinical isolates and results were compared to results of the solid-phase enzyme immunoassay typing method. These isolates represented the most prevalent genital serovars (D, E, F, K, I and J) in the local sexually transmitted diseases clinic population. For specimens containing 1 serovar, the results of the two methods were similar for 42 samples and discordant for 1 sample. In addition, two samples showed evidence of mixed infection with two serovars as identified by both methods. Five additional specimens contained two serovars, as shown by one or both methods. In all five such specimens, the two typing methods agreed on at least one of the two serovars. For both single and multiple serovar specimens, there was concordance between the two typing methods for 16/17 E serovars, 8/9 D serovars, 8/8 F serovars, 7/7 I serovars, 7/7 J serovars, 5/8 K serovars, and 0/2 G serovars. This rapid and economic method can detect mixed infections, may circumvent the need to culture chlamydial organisms for typing, and should be a useful addition to chlamydia typing technology.

Original languageEnglish (US)
Pages (from-to)303-308
Number of pages6
JournalSexually Transmitted Diseases
Volume19
Issue number6
StatePublished - 1992

Fingerprint

Chlamydia trachomatis
DNA Restriction Enzymes
Digestion
Polymerase Chain Reaction
Genes
Serogroup
Coinfection
Restriction Fragment Length Polymorphisms
Membrane Proteins
Chlamydia
Sexually Transmitted Diseases
Immunoenzyme Techniques
Silver
Base Pairing

ASJC Scopus subject areas

  • Dermatology
  • Infectious Diseases
  • Microbiology (medical)
  • Public Health, Environmental and Occupational Health

Cite this

Gene typing of chlamydia trachomatis by polymerase chain reaction and restriction endonuclease digestion. / Gaydos, Charlotte A; Bobo, Linda; Welsh, Laura; Hook, Edward W.; Viscidi, Raphael P; Quinn, Thomas C.

In: Sexually Transmitted Diseases, Vol. 19, No. 6, 1992, p. 303-308.

Research output: Contribution to journalArticle

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