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Qualitative and Quantitative Detection of Chlamydophila pneumoniae DNA in Cerebrospinal Fluid from Multiple Sclerosis Patients and Controls

By Yi-Wei Tang, Subramaniam Sriram, Haijing Li, Song-yi Yao, Shufang Meng, William M. Mitchell and Charles W. Stratton


A standardized molecular test for the detection of Chlamydophila pneumoniae DNA in cerebrospinal fluid (CSF) would assist the further assessment of the association of C. pneumoniae with multiple sclerosis (MS). We developed and validated a qualitative colorimetric microtiter plate-based PCR assay (PCR-EIA) and a real-time quantitative PCR assay (TaqMan) for detection of C. pneumoniae DNA in CSF specimens from MS patients and controls. Compared to a touchdown nested-PCR assay, the sensitivity, specificity, and concordance of the PCR-EIA assay were 88.5%, 93.2%, and 90.5%, respectively, on a total of 137 CSF specimens. PCR-EIA presented a significantly higher sensitivity in MS patients (p = 0.008) and a higher specificity in other neurological diseases (p = 0.018). Test reproducibility of the PCR-EIA assay was statistically related to the volumes of extract DNA included in the test (p = 0.033); a high volume, which was equivalent to 100 µl of CSF per reaction, yielded a concordance of 96.8% between two medical technologists running the test at different times. The TaqMan quantitative PCR assay detected 26 of 63 (41.3%) of positive CSF specimens that tested positive by both PCR-EIA and nested-PCR qualitative assays. None of the CSF specimens that were negative by the two qualitative PCR methods were detected by the TaqMan quantitative PCR. The PCR-EIA assay detected a minimum of 25 copies/ml C. pneumoniae DNA in plasmid-spiked CSF, which was at least 10 times more sensitive than TaqMan. These data indicated that the PCR-EIA assay possessed a sensitivity that was equal to the nested-PCR procedures for the detection of C. pneumoniae DNA in CSF. The TaqMan system may not be sensitive enough for diagnostic purposes due to the low C. pneumoniae copies existing in the majority of CSF specimens from MS patients

Topics: Research Article
Publisher: Public Library of Science
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Provided by: PubMed Central
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    1. (1990). A new respiratory tract pathogen: Chlamydia pneumoniae strain TWAR.
    2. (1999). Chlamydia pneumoniae infection of the central nervous system in multiple sclerosis.
    3. (1998). Comparative evaluation of colometric microtiter plate systems for detection of herpes simplex virus in cerebrospinal fluid.
    4. (2004). Comparison of five PCR assays for detecting Chlamydia pneumoniae DNA.
    5. (1994). Detection of Chlamydia pneumoniae in clinical specimens by polymerase chain reaction using nested primers.
    6. (1985). Evolution of the functional human beta-actin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes.
    7. (2005). Methodological aspects affecting the infectivity of Chlamydia pneumoniae in cell cultures in vitro.
    8. (2002). Molecular approaches to detecting herpes simplex and enteroviruses in the central nervous system.
    9. (1999). Molecular diagnosis of Chlamydia pneumoniae infection.
    10. (2006). Multiple sclerosis: an infectious syndrome involving Chlamydophila pneumoniae.
    11. (2002). PCR in diagnosis of infection: detection of bacteria in cerebrospinal fluids.
    12. (2001). PCR-based method for isolation and detection of Chlamdyia pneumoniae DNA in cerebrospinal fluids.
    13. (2000). Petrich A
    14. (1997). Rapid diagnosis of respiratory Chlamdyia pneumoniae infection by nested touchdown polymerase chain reaction compared with culture and antigen detection by EIA.
    15. (1991). Sequence analysis of the major outer membrane protein gene of Chlamydia pneumoniae.
    16. (2001). Standardizing Chlamydia pneumoniae assays: recommendations from the Centers for Diseases Control and Prevention (USA) and the Laboratory Centre for Disease Control (Canada).
    17. (2008). Under the microscope: focus on Chlamydia pneumoniae infection and multiple sclerosis.

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