title: Comparison of two different PCR detection methods. Application to the diagnosis of pulmonary tuberculosis.
abstract: The objectives are to assess the influence of the detection of the amplified DNA fragment on the sensitivity and specificity of the polymerase chain reaction (PCR). One hundred seventy-five sputum samples from 123 patients were processed. Sixty samples were taken from 60 subjects without tuberculosis, and the rest were taken from subjects with tuberculosis confirmed by culture. A fragment of the IS6110 sequence of Mycobacterium tuberculosis, which was detected using two different methods, was amplified. The detection methods used were a digoxigenin-labeled specific probe and chemiluminescent development and reamplification (nested PCR) combined with agarose gel electrophoresis. Sensitivity with probe detection was 75.65% and specificity 100%. Using the nested PCR technique, sensitivity rose to 93.04%, but specificity decreased to 96.6%. PCR is a quick and adequate way to diagnose pulmonary tuberculosis in cases where staining is negative yet there is a clinical suspicion of tuberculosis, even though a standardization process and large scale evaluation are still needed to determine its true usefulness.
authors.affiliation: Department of Microbiology, General University Hospital of Elche, University of Alicante, Spain.
authors.forename: J C
authors.initials: JC
authors.lastname: Rodríguez
authors.forename: E
authors.initials: E
authors.lastname: Fuentes
authors.forename: G
authors.initials: G
authors.lastname: Royo
mesh_terms: D004251:DNA Transposable Elements / Q000235:genetics; D005188:False Negative Reactions; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis*
chemical_list: D004251:DNA Transposable Elements