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Journal of Clinical Microbiology, November 2002, p. 4230-4234, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.4230-4234.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of the Causative Organism of Tuberculous Lymphadenitis in Ethiopia by PCR

Dawit Kidane,1 Joseph O. Olobo,1,2 Abebe Habte,1 Yohannes Negesse,1 Abraham Aseffa,1 Getahun Abate,1 Mohammed A. Yassin,3 Kiflu Bereda,4 and Morten Harboe1,5*

Armauer Hansen Research Institute (AHRI), Addis Ababa,1 Southern Region Health Bureau, Awassa,3 Butajira Health Center, Southern Region, Butajira District, Ethiopia,4 Makerere University, Kampala, Uganda,2 Institute of Immunology, The National Hospital, N-0027 Oslo, Norway5

Received 30 April 2002/ Returned for modification 17 August 2002/ Accepted 29 August 2002

Tuberculous lymphadenitis (TBLN) is a common form of extrapulmonary tuberculosis with multiple differential diagnoses. Demonstration of the etiologic agent by smear microscopy or culture of fine needle aspirate (FNA) specimens is often unsuccessful. FNA specimens from 40 patients presenting at a rural health center in South Ethiopia and diagnosed as positive for TBLN on the basis of clinical and cytological criteria were analyzed for mycobacterial DNA by PCR. Thirty (75%) had cervical lymphadenitis and 11 (27.5%) were seropositive for human immunodeficiency virus (HIV). Three primer sets were initially used to identify the causative agent at the genus (antigen 85 complex), complex (IS6110 insertion sequence), and species (pncA gene and allelic variation) levels. Among the forty TBLN cases, 35 (87.5%) were positive by PCR at the genus and complex levels. Based on PCR for detection of allelic variation at position 169, 24 (68.6%) of the 35 were positive for Mycobacterium tuberculosis and 6 (17.1%) were positive for M. bovis. These six were positive in additional PCR assays using the JB21-JB22 primer set, which is highly specific for M. bovis. Five (14.1%) showed amplification for both M. tuberculosis and M. bovis with the allele-specific primer set. Cooccurrence of pyrazinamide (PZA)-sensitive and -resistant M. tuberculosis in those five cases was indicated, since all were negative in assays with the JB21-JB22 primer set. This feature was seen in 3 of 11 HIV-positive and 2 of 29 HIV-negative individuals (P < 0.001). Conclusion: among 35 PCR-positive cases of TBLN from southern Ethiopia, 29 (82.9%) were caused by M. tuberculosis and six (17.1%) were caused by M. bovis.

* Corresponding author: Mailing address: Armauer Hansen Research Institute, P.O. Box 1005, Addis Ababa, Ethiopia. Phone: 251 1 71 02 88. Fax: 251 1 71 13 90. E-mail: ahri{at}telecom.net.et.

Journal of Clinical Microbiology, November 2002, p. 4230-4234, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.4230-4234.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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