title: PCR-based method for isolation and detection of Chlamydia pneumoniae DNA in cerebrospinal fluids.
abstract: Since current studies indicate the possible involvement of Chlamydia pneumoniae in the pathogenesis of multiple sclerosis (MS), demonstration of C. pneumoniae in the cerebrospinal fluid (CSF) of patients with MS is highly desirable. However, there is controversy concerning the detection of C. pneumoniae in CSFs from MS patients due to the lack of a standard protocol for extraction and detection of C. pneumoniae DNA. In this regard, we attempted to establish a highly effective extraction protocol for C. pneumoniae DNA from CSFs utilizing a commercial kit and a PCR detection method. The extraction and PCR detection protocol established in this study succeeded in detecting as few as 20 C. pneumoniae organisms in 200 microl of mock CSF. The use of this protocol to detect C. pneumoniae DNA in CSFs revealed that 68% of CSF samples obtained from patients with MS were positive (11 out of 16 samples) for chlamydia DNA. Thus, the protocol established here is sensitive enough to detect chlamydia DNA from CSFs and can be used by other laboratories for evaluation of the presence of chlamydiae in CSFs because the protocol is based on the use of a commercial kit.
authors.affiliation: Department of Medical Microbiology and Immunology, University of South Florida College of Medicine, Tampa, Florida 33612, USA.
authors.forename: H
authors.initials: H
authors.lastname: Ikejima
authors.forename: S
authors.initials: S
authors.lastname: Haranaga
authors.forename: H
authors.initials: H
authors.lastname: Takemura
authors.forename: T
authors.initials: T
authors.lastname: Kamo
authors.forename: Y
authors.initials: Y
authors.lastname: Takahashi
authors.forename: H
authors.initials: H
authors.lastname: Friedman
authors.forename: Y
authors.initials: Y
authors.lastname: Yamamoto
mesh_terms: D023521:Chlamydophila Infections / Q000134:cerebrospinal fluid / Q000382:microbiology; D016993:Chlamydophila pneumoniae / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid*; D006801:Humans; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid / Q000209:etiology / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial