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Erschienen in: Zeitschrift für Rheumatologie 9/2015

01.11.2015 | Originalien

Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice

verfasst von: J. Freise, I. Bernau, S. Meier, H. Zeidler, Prof. Dr. J.G. Kuipers

Erschienen in: Zeitschrift für Rheumatologie | Ausgabe 9/2015

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Abstract

Aim

No standardized polymerase chain reaction (PCR) assay is available for detection of Chlamydia trachomatis (C. tr.) in synovial fluid (SF) for diagnostic use in clinical practice. This study tested the performance of two optimized molecular biology methods, to determine which is best suited for detecting C. tr. in SF clinical samples from patients with various rheumatologic diseases.

Methods

Two DNA extraction methods, i.e., (1) alkaline lysis and (2) QIAEX II Gel Extraction Kit® + cetyltrimethylammonium bromide (CTAB; Qiagen, Hilden, Germany), and C. tr.-omp1-152 bp PCR were tested in SF samples from a total of 329 patients with the following diagnoses: reactive arthritis (ReA; n = 10, 4 patients had posturethritic ReA), undifferentiated arthritis (UA; n = 66), rheumatoid arthritis (RA; n = 169), psoriatic arthritis (PSA; n = 12), and osteoarthritis (OA) n = 72.

Results

In SF samples, C. tr.-omp1-152 bp PCR in combination with alkaline lysis DNA extraction allowed detection of more C. tr.-positive samples: 3/10 (30 %) ReA patients (all with posturethritic ReA) and 20/66 (38 %) UA patients were positive, compared to the 0/10 (0 %) patients with ReA and 1/66 (2 %) with UA detected using the QIAEX II Gel Extraction Kit® + CTAB. Moreover, 2/12 (17 %) SF samples from PSA patients tested positive with alkaline lysis. All samples from patients with OA and RA tested negative.

Conclusion

Alkaline lysis in combination with C. tr.-omp1-152 bp PCR emerged as the most sensitive method for identification of C. tr. in clinical SF samples.
Literatur
1.
Zurück zum Zitat Bas S, Griffais R, Kvien TK et al (1995) Amplification on plasmid and chromosome Chlamydia DNA in synovial fluid of patients with reactive arthritis and undifferentiated seronegative oligoarthropathies. Arthritis Rheum 38:1005–1013CrossRefPubMed Bas S, Griffais R, Kvien TK et al (1995) Amplification on plasmid and chromosome Chlamydia DNA in synovial fluid of patients with reactive arthritis and undifferentiated seronegative oligoarthropathies. Arthritis Rheum 38:1005–1013CrossRefPubMed
2.
Zurück zum Zitat Beutler AM, Whittum-Hudson JA, Nanagara R et al (1994) Intracellular location of inapparently infecting Chlamydia in synovial tissue from patients with Reiter’s syndrome. Immunol Res 13:163–171CrossRefPubMed Beutler AM, Whittum-Hudson JA, Nanagara R et al (1994) Intracellular location of inapparently infecting Chlamydia in synovial tissue from patients with Reiter’s syndrome. Immunol Res 13:163–171CrossRefPubMed
4.
Zurück zum Zitat Bobo L, Coutlee F, Yolken R et al (1990) Diagnosis of Chlamydia trachomatis cervical infection by detection of amplified DNA with an enzyme immunoassay. J Clin Microbiol 28:1968–1973PubMedCentralPubMed Bobo L, Coutlee F, Yolken R et al (1990) Diagnosis of Chlamydia trachomatis cervical infection by detection of amplified DNA with an enzyme immunoassay. J Clin Microbiol 28:1968–1973PubMedCentralPubMed
5.
Zurück zum Zitat Freise J, Bernau I, Meier S et al (2009) Detection of Chlamydia trachomatis-DNA in synovial fluid: evaluation of the sensitivity of different DNA extraction methods and amplification systems. Arthritis Res Ther 11:R175PubMedCentralCrossRefPubMed Freise J, Bernau I, Meier S et al (2009) Detection of Chlamydia trachomatis-DNA in synovial fluid: evaluation of the sensitivity of different DNA extraction methods and amplification systems. Arthritis Res Ther 11:R175PubMedCentralCrossRefPubMed
6.
Zurück zum Zitat Freise J, Gerard HC, Bunke T et al (2001) Optimised sample DNA preparation for detection of Chlamydia trachomatis in synovial tissue by polymerase chain reaction and ligase chain reaction. Ann Rheum Dis 60:140–145PubMedCentralCrossRefPubMed Freise J, Gerard HC, Bunke T et al (2001) Optimised sample DNA preparation for detection of Chlamydia trachomatis in synovial tissue by polymerase chain reaction and ligase chain reaction. Ann Rheum Dis 60:140–145PubMedCentralCrossRefPubMed
7.
Zurück zum Zitat Gerard HC, Branigan PJ, Schumacher HR Jr, Hudson AP (1998) Synovial Chlamydia trachomatis in patients with reactive arthritis/Reiter’s syndrome are viable but show aberrant gene expression. J Rheumatol 25:734–742PubMed Gerard HC, Branigan PJ, Schumacher HR Jr, Hudson AP (1998) Synovial Chlamydia trachomatis in patients with reactive arthritis/Reiter’s syndrome are viable but show aberrant gene expression. J Rheumatol 25:734–742PubMed
8.
Zurück zum Zitat Kuipers JG, Andresen J, Köhler L et al (2002) Evaluation of amplicor chlamydia PCR and LCX chlamydia LCR to detect Chlamydia trachomatis in synovial fluid. Clin Exp Rheumatol 20:185–192PubMed Kuipers JG, Andresen J, Köhler L et al (2002) Evaluation of amplicor chlamydia PCR and LCX chlamydia LCR to detect Chlamydia trachomatis in synovial fluid. Clin Exp Rheumatol 20:185–192PubMed
9.
Zurück zum Zitat Kuipers JG, Nietfeld L, Dreses-werringloer U et al (1999) Optimized sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction. Ann Rheum Dis 58:103–108PubMedCentralCrossRefPubMed Kuipers JG, Nietfeld L, Dreses-werringloer U et al (1999) Optimized sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction. Ann Rheum Dis 58:103–108PubMedCentralCrossRefPubMed
10.
Zurück zum Zitat Kuipers JG, Scharmann K, Wollenhaupt J et al (1995) Sensitivities of PCR, MicroTrak, ChlamydiaEIA, IDEIA, and PACE 2 for purified Chlamydia trachomatis elementary bodies in urine, peripheral blood, peripheral blood leukocytes, and synovial fluid. J Clin Microbiol 33:3186–3190PubMedCentralPubMed Kuipers JG, Scharmann K, Wollenhaupt J et al (1995) Sensitivities of PCR, MicroTrak, ChlamydiaEIA, IDEIA, and PACE 2 for purified Chlamydia trachomatis elementary bodies in urine, peripheral blood, peripheral blood leukocytes, and synovial fluid. J Clin Microbiol 33:3186–3190PubMedCentralPubMed
11.
Zurück zum Zitat Kuipers JG, Sibilia J, Bas S et al (2009) Reactive and undifferentiated arthritis in North Africa: use of PCR for detection of Chlamydia trachomatis. Clin Rheumatol 28:11–16CrossRefPubMed Kuipers JG, Sibilia J, Bas S et al (2009) Reactive and undifferentiated arthritis in North Africa: use of PCR for detection of Chlamydia trachomatis. Clin Rheumatol 28:11–16CrossRefPubMed
12.
Zurück zum Zitat Møller JK, Pedersen LN, Persson K (2010) Comparison of the Abbott RealTime CT new formulation assay with two other commercial assays for detection of wild-type and new variant strains of Chlamydia trachomatis. J Clin Microbiol 48:440–443PubMedCentralCrossRefPubMed Møller JK, Pedersen LN, Persson K (2010) Comparison of the Abbott RealTime CT new formulation assay with two other commercial assays for detection of wild-type and new variant strains of Chlamydia trachomatis. J Clin Microbiol 48:440–443PubMedCentralCrossRefPubMed
13.
Zurück zum Zitat Olmez N, Wang GF, Li Y et al (2001) Chlamydial nucleic acids in synovium in osteoarthritis: what are the implications? J Rheumatol 28:1874–1880PubMed Olmez N, Wang GF, Li Y et al (2001) Chlamydial nucleic acids in synovium in osteoarthritis: what are the implications? J Rheumatol 28:1874–1880PubMed
14.
Zurück zum Zitat Priem S, Rittig MG, Kamradt T et al (1997) An optimized PCR leads to rapid and highly sensitive detection of Borrelia burgdorferi in patients with Lyme borreliosis. J Clin Microbiol 35:685–690PubMedCentralPubMed Priem S, Rittig MG, Kamradt T et al (1997) An optimized PCR leads to rapid and highly sensitive detection of Borrelia burgdorferi in patients with Lyme borreliosis. J Clin Microbiol 35:685–690PubMedCentralPubMed
15.
Zurück zum Zitat Schnarr S, Putschky N, Jendro MC et al (2001) Chlamydia and Borrelia DNA in synovial fluid of patients with early undifferentiated oligoarthritis: results of a prospective study. Arthritis Rheum 44:2679–2685CrossRefPubMed Schnarr S, Putschky N, Jendro MC et al (2001) Chlamydia and Borrelia DNA in synovial fluid of patients with early undifferentiated oligoarthritis: results of a prospective study. Arthritis Rheum 44:2679–2685CrossRefPubMed
16.
Zurück zum Zitat Siala M, Gdoura R, Younes M et al (2009) Detection and frequency of Chlamydia trachomatis DNA in synovial samples from Tunisian patients with reactive arthritis and undifferentiated oligoarthritis. FEMS Immunol Med Microbiol 55:178–186CrossRefPubMed Siala M, Gdoura R, Younes M et al (2009) Detection and frequency of Chlamydia trachomatis DNA in synovial samples from Tunisian patients with reactive arthritis and undifferentiated oligoarthritis. FEMS Immunol Med Microbiol 55:178–186CrossRefPubMed
17.
Zurück zum Zitat Silveira LH, Gutierrez F, Scopelitis E et al (1993) Chlamydia- induced arthritis. Rheum Dis Clin North Am 19:351–363PubMed Silveira LH, Gutierrez F, Scopelitis E et al (1993) Chlamydia- induced arthritis. Rheum Dis Clin North Am 19:351–363PubMed
18.
Zurück zum Zitat Taylor-Robinson D, Gilroy CB, Thomas BJ, Keat AC (1992) Detection of Chlamydia trachomatis DNA in joints of reactive arthritis patients by polymerase chain reaction. Lancet 340:81–82CrossRefPubMed Taylor-Robinson D, Gilroy CB, Thomas BJ, Keat AC (1992) Detection of Chlamydia trachomatis DNA in joints of reactive arthritis patients by polymerase chain reaction. Lancet 340:81–82CrossRefPubMed
19.
Zurück zum Zitat Wilkinson NZ, Kingsley GH, Sieper J et al (1998) Lack of correlation between the detection of Chlamydia trachomatis DNA in synovial fluid from patients with a range of rheumatic diseases and the presence of an antichlamydial immune response. Arthritis Rheum 41:845–854CrossRefPubMed Wilkinson NZ, Kingsley GH, Sieper J et al (1998) Lack of correlation between the detection of Chlamydia trachomatis DNA in synovial fluid from patients with a range of rheumatic diseases and the presence of an antichlamydial immune response. Arthritis Rheum 41:845–854CrossRefPubMed
20.
Zurück zum Zitat Wollenhaupt J, Schnarr S, Kuipers JG (1998) Bacterial antigens in reactive arthritis and spondarthritis. Rational use of laboratory testing in diagnosis and follow-up. Baillieres Clin Rheumatol 12:627–647CrossRefPubMed Wollenhaupt J, Schnarr S, Kuipers JG (1998) Bacterial antigens in reactive arthritis and spondarthritis. Rational use of laboratory testing in diagnosis and follow-up. Baillieres Clin Rheumatol 12:627–647CrossRefPubMed
21.
Zurück zum Zitat Zeidler H, Kuipers JG, Köhler L (2004) Chlamydia- induced arthritis. Curr Op Rheumatol 16:380–392CrossRef Zeidler H, Kuipers JG, Köhler L (2004) Chlamydia- induced arthritis. Curr Op Rheumatol 16:380–392CrossRef
Metadaten
Titel
Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice
verfasst von
J. Freise
I. Bernau
S. Meier
H. Zeidler
Prof. Dr. J.G. Kuipers
Publikationsdatum
01.11.2015
Verlag
Springer Berlin Heidelberg
Erschienen in
Zeitschrift für Rheumatologie / Ausgabe 9/2015
Print ISSN: 0340-1855
Elektronische ISSN: 1435-1250
DOI
https://doi.org/10.1007/s00393-015-1589-y

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