Tuberculosis is still one of the most important cause of mortality and morbidity in many countries and there is a need for new methods for accurate and rapid diagnosis of tuberculosis. To determine the sensitivity and specificity of polymerase chain reaction (PCR) method, we have evaluated Mycobacterium tuberculosis DNA in peripheral blood samples with PCR technique in adult patients with human immunodeficiency virus (HIV)-negative and new cases of smear-positive pulmonary tuberculosis. We investigated the relationship between characteristic of the patients, radiological extension of the disease, sputum smear grade, presence of cavity, body-mass index (BMI), serum albumin level, total delay time and PCR positivity. Forty patients (33 male and 7 female; mean age 37.8±14.1) and 20 healthy control subjects (13 male and 7 female; mean age 35.6±7.3) were enrolled in this study. PCR was positive in 16 of 40 (40%) patients with pulmonary tuberculosis and negative in 24 of 40 (60%). None of the healthy controls had positive PCR results. The overall sensitivity, specificity and accuracy of the PCR assay was 40,100 and 60%, respectively. We found the positive correlation between PCR positivity and sputum smear grade (r= 0.46, P=0.003), radiological extension of the disease (r= 0.69, P=0.001), presence of cavity (r= 0.90, P=0.001). We conclude that the detection of M. tuberculosis DNA from peripheral blood by PCR technique is useful for the rapid diagnosis of tuberculosis patients with HIV-negative. Hematogenous dissemination was important in tuberculosis patients and peripheral blood samples were suitable and easy materials. However, standardization of the PCR method must be ensured for the diagnosis of tuberculosis. © 2003 Elsevier Science Ltd. All rights reserved.