title: Comparison of three methods of DNA extraction in endocervical specimens for Chlamydia trachomatis infection by spectrophotometry, agarose gel, and PCR.
abstract: Chlamydia trachomatis is the major cause of sexually transmitted disease in the world. The aim of this study was to determine the best method of DNA extraction for detecting C. trachomatis by polymerase chain reaction (PCR) in sexually active women (n = 80) attending Shahid Beheshti Hospital in Isfahan, Iran. Endocervical swabs were collected from 80 women, 22 of whom were asymptomatic and 58 symptomatic. Three different DNA extraction methods were used in this study (phenol-chlorophorm, proteinase K, and boiling). DNA yield was evaluated by spectrophotometry, agarose gel, and PCR. The internal control was assayed by beta-globin primers (PCO4, GH20). The DNA cryptic plasmid was selected as the target for C. trachomatis and samples were examined by PCR using specific KL1 and KL2 primers. It was shown that DNA extraction by boiling was the most sensitive with the highest yield of DNA. Of the 80 samples, 17 (21.25%) showed positivity for C. trachomatis by PCR. The highest rate of C. trachomatis infection was found in the group aged between 35 and 45 years old and those who used withdrawal or an intrauterine device as methods of contraception. It was demonstrated that DNA extraction by boiling was the least expensive and a very rapid method that gave the highest DNA yield. The infection rate in the sexually active women, including symptomatic and asymptomatic, was 21.25%, with a presumably high prevalence compared with other studies done in this field.
authors.affiliation: Section of Microbiology, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran.
authors.forename: Anahita
authors.initials: A
authors.lastname: Jenab
authors.forename: Rasoul
authors.initials: R
authors.lastname: Roghanian
authors.forename: Naser
authors.initials: N
authors.lastname: Golbang
authors.forename: Pouran
authors.initials: P
authors.lastname: Golbang
authors.forename: Leili
authors.initials: L
authors.lastname: Chamani-Tabriz
mesh_terms: D000328:Adult; D002584:Cervix Uteri / Q000378:metabolism / Q000473:pathology; D002690:Chlamydia Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000235:genetics* / Q000472:pathogenicity; D003267:Contraception; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D016133:Polymerase Chain Reaction; D015995:Prevalence; D052616:Solid Phase Extraction / Q000379:methods; D013053:Spectrophotometry
chemical_list: D004269:DNA, Bacterial