Detection of Mycobacterium Tuberculosis DNA from peripheral blood in patients with HIV-seronegative and new cases of smear-positive pulmonary tuberculosis by polymerase chain reaction

Abstract

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.

Evaluating bacterial pathogen DNA preservation in museum osteological collections

“…55-75%: Haas et al 2000;Zink et al 2001a;Fletcher et al 2003;Zink et al 2003). These latter data compare favourably with published rates of detection using simple PCR-based systems on modern, diagnosed, clinical samples, which are around 80% (Portillo-Gomez et al 2000;Van der Spoel van Dijk et al 2000;Mitarai et al 2001;Narayanan et al 2001;Alfonso et al 2002;Yee et al 2002;Leung et al 2003;Cheng et al 2004), and higher than detection rates for blood (40%: Taci et al 2003), urine (56%: Kafwabulula et al 2002), and host DNA in studies of animal bones from temperate archaeological sites (around 10-20%: Haynes et al 2002;Edwards et al 2004). The high frequency of amplification success from archaeological samples has been attributed to the enhanced stability of the M. tuberculosis cell wall (Donoghue et al 2004).…”

Tuberculous otitis media: a case presentation and review of the literature

“…Gene amplification techniques such as PCR can be useful 33. If the cause of suppurative infection of the middle ear is still undiagnosed, then operative biopsy may be required for diagnosis 30 .…”

Exploring Alternative Biomaterials for Diagnosis of Pulmonary Tuberculosis in HIV-Negative Patients by Use of the GeneXpert MTB/RIF Assay

“…tuberculosis has been detected in blood samples from TB patients with variable degrees of sensitivity (10-12, 15, 16, 38, 39). Most TB patients with positive blood tests have been HIV positive (12), and the highest sensitivities reported for HIV-negative patients were 43.8% and 40% (11). However, many of these prior studies were poorly controlled, and the specificity of the assay used was often uncertain.…”

“…Blood and urine samples are also relatively simple to collect from patients. A number of reports have detected DNA from Mycobacterium tuberculosis in blood and urine samples from HIV-positive and -negative patients with pulmonary TB (10)(11)(12)(13)(14)(15)(16); however, results of individual studies have been quite variable (10-12, 14, 15).…”