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Journal of Clinical Microbiology, July 2004, p. 3036-3040, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3036-3040.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Improved Sensitivity of Nucleic Acid Amplification for Rapid Diagnosis of Tuberculous Meningitis

Isik Somuncu Johansen,1* Bettina Lundgren,2 Fehmi Tabak,3 Björn Petrini,4 Salih Hosoglu,5 Nese Saltoglu,6 and Vibeke Østergaard Thomsen1

International Reference Laboratory of Mycobacteriology, Statens Serum Institut,1 Department of Clinical Microbiology, Hvidovre University Hospital, Copenhagen, Denmark,2 Department of Infectious Diseases and Clinical Microbiology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul,3 Department of Infectious Diseases, Dicle University Hospital, Diyarbakir,5 Department of Infectious Diseases and Clinical Microbiology, Faculty of Medicine, Cukurova University, Adana, Turkey,6 Department of Clinical Microbiology, Karolinska Institute and Hospital, Stockholm, Sweden4

Received 29 December 2003/ Returned for modification 4 March 2004/ Accepted 28 March 2004

Early diagnosis of tuberculous meningitis (TBM) is essential for a positive outcome; but present microbiological diagnostic techniques are insensitive, slow, or laborious. We evaluated the standard BDProbeTec ET strand displacement amplification method (the standard ProbeTec method) for the detection of Mycobacterium tuberculosis complex organisms in parallel with the ProbeTec method with a modified pretreatment procedure with 101 prospectively collected cerebrospinal fluid specimens from 94 patients with suspected TBM. By the modified method, the sample-washing step was omitted. A definitive diagnosis was attained by culture. Thirteen specimens from 12 patients were culture positive for M. tuberculosis complex organisms; three specimens (23%) were microscopy positive for acid-fast bacilli. Among the culture-positive specimens, the standard ProbeTec method was positive for 8 (61.5%) and the modified assay was positive for 10 (76.9%). The overall specificity by both procedures was 98.8% compared to the results of culture. After discrepancy analysis, conducted by reviewing the patients' previous laboratory data, the specificity increased to 100%. If the cutoff value for respiratory specimens was adjusted from the recommended value of 3,400 to 1,000, the sensitivity of the modified procedure increased to 84.7%, with unchanged specificity. Results were obtained in 3 to 4 h. The new pretreatment procedure with the ProbeTec assay described here provides a rapid, simple, and sensitive tool for the diagnosis of TBM.

* Corresponding author. Mailing address: International Reference Laboratory of Mycobacteriology, Statens Serum Institut, 5 Artillerivej, 2300 Copenhagen S, Denmark. Phone: 4532683703. Fax: 4532683871. E-mail: isj{at}ssi.dk.

Journal of Clinical Microbiology, July 2004, p. 3036-3040, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3036-3040.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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