MICROBIOLOGY LETTERS zyxwvutsrqponmlkjihg ELSEVIER FEMS Microbiology Letters 130 (1995) 205-210 Detection by multiplex polymerase chain reaction and typing of Chlamydia trachomatis isolates Marcello Valassina ap * , Maria Grazia Cusi a, Daniele Corsaro a, Carlo Buffi b, Giovanna Piazzesi b, Pier Egisto Valensin a a Department of Molecular Biology, Microbiology Section, Vniuersiry of Siena, via Laterina 8, 53100 Siena, Italy b Gynecological and Obstetric Department, Burresi Hospital, Poggibonsi (Siena), Italy Received 10 April 1995; revised 29 May 1995; accepted 30 May 1995 zyxwvutsrqponmlkjihgfedcbaZYXWVU Abstract The multiplex polymerase chain reaction (PCR) was applied for the detection of the Chlumydia trachomatis chromosome and plasmid. The multiplex PCR demonstrated a sensitivity of 0.8 fg of chlamydial DNA, corresponding to the detection of about 5 copies of the plasmid. Analysis of 195 genital specimens collected randomly from a female population, showed that the multiplex PCR is more sensitive and rapid than culturing for detecting Chlamydia trachomatis. Moreover, sequencing of the II variable domain of the ompl gene, directly from DNA of the clinical specimens, appears to be a simple and rapid method for determining serovar isolates. Keywords: Chlamydia spp.; Genotype; Polymerase chain reaction 1. Introduction Chlamydia trachomatis is an ubiquitous microor- ganism and one of the most common agents of sexually transmitted diseases: urethritis, epididymi- tis, prostatitis in men, and cervicitis, endometritis, and salpingitis in women. Without antibiotic inter- vention, long-term chlamydial infections of the fe- male genital tract may result in chronic manifesta- tions eventually complicated with tubal infertility and ectopic pregnancy [l]. The requirement for an * Corresponding author. Tel: + 39-577-280903; Fax: + 39-577- 420. early diagnosis of chlamydia infection can be satis- fied by using the polymerase chain reaction (PCR), a technique that can compete with the cell culture method both for specificity and sensitivity [2]. Both restriction fragment length polymorphism (RFLP) analysis [3] and direct sequencing [4], have been used for typing chlamydial isolates. The hyper- variable regions of the ompl gene coding for the major outer membrane protein (MOMP) have been the main targets of these typing systems [51. Simultaneous amplification of the chlamydial plasmid and chromosomal DNA was used to support laboratory diagnosis of C. trachomatis. Rapid geno- typing of isolates was done by direct sequencing of PCR products. Data on the occurrence of C. tra- chomatis in a random female population are shown. 037%1097/95/$09.50 0 1995 Federation of European Microbiological Societies. All rights reserved SSDI 0378-1097(95)00207-3 Downloaded from https://academic.oup.com/femsle/article-abstract/130/2-3/205/484131 by guest on 21 May 2020