Optimised sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction

doi:10.1136/ard.58.2.103

Annals of the Rheumatic Diseases 1999;58:103-108; doi:10.1136/ard.58.2.103

Ann Rheum Dis 1999;58:103-108 ( February )

Extended reports

Optimised sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction Jens G Kuipers,^a Lars Nietfeld,^a Ute Dreses-Werringloer,^a Lars Koehler,^a Juergen Wollenhaupt,^a Henning Zeidler,^a Michael Hammer^b

^a Division of Rheumatology, Department of Internal Medicine and Dermatology, Hannover Medical School, Hannover, Germany, ^b Department of Rheumatology, Nordwestdeutsches Rheumazentrum, St Josef-Stift, Sendenhorst, Germany

Correspondence to: Dr J Kuipers, Division of Rheumatology, Hannover Medical School, 30623 Hannover, Germany.

Accepted for publication 13 October 1998

OBJECTIVE $---$ To optimise sample preparation of synovial fluid for Chlamydia trachomatis (CT) specific polymerase chain reaction(PCR).
METHODS $---$ Serial dilutions of purified CT elementary bodies in synovial fluid were prepared. The synovial fluid pellet was processedby eight different methods of sample preparation. Then sampleswere analysed by CT specific PCR. The sensitivity of PCR was thebasis of ranking of the eight differentmethods.
RESULTS $---$ Highest sensitivity was achieved by methods including an additional step of DNA isolation. Additional extraction of proteinand polysaccharides by cetyltrimethylammonium bromide (CTAB) increasedsensitivity. Addition of hyaluronidase did not increase sensitivityof QIAEX-DNA extraction but was necessary, however, before phenol-chloroform-DNAextraction.
CONCLUSIONS $---$ The method of synovial fluid sample preparation significantly influences the sensitivity of subsequent PCR. Additional DNAisolation and extraction of PCR inhibitors by CTAB led to highersensitivity.

Keywords: Chlamydia trachomatis; polymerase chain reaction; synovial fluid

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