Finally, these false-positive cultures lead to an overestimation of the incidence and prevalence of tuberculosis in humans [10]. A definitive demonstration of cross-contamination can be derived from precise molecular analyses of M. tuberculosis isolates. M. tuberculosis
isolates harbouring PLK inhibitor identical genotypes are regarded as clones and are thus epidemiologically linked [11]. The most widely used technique for determining the genotype of M. tuberculosis is a technique known as IS6110-restriction fragment length polymorphism (RFLP) analysis. RFLP analysis requires a large amount of biological material and, thus, poses a risk to laboratory workers due to the harmful nature of this pathogen. Moreover, the latter method requires a substantial amount of time due to the fastidious nature of M. www.selleckchem.com/products/Trichostatin-A.html tuberculosis [12]. More importantly from, a strictly technical perspective, IS6110-RFLP analysis does a poor job of indicating the presence of M. tuberculosis when these organisms contain only a few copies of the IS6110 sequence [13]. Recently, the variable number tandem repeat (VNTR) PCR-based technique and the mycobacterial interspersed GS-4997 in vitro repetitive unit (MIRU) [14]
technique have proven to be reliable methods for the resolution of cross-contamination events [15, 16]. We herein report the application of a new PCR-sequencing-based genotyping method, known as multispacer sequence typing (MST)[17], for determining whether specimens have been cross-contaminated with M. tuberculosis in the laboratory. Case report A 60-year-old man was admitted for an examination to determine whether he had interstitial pneumonia.
The patient had been previously Interleukin-2 receptor hospitalised for two weeks at a different location with symptoms that included shortness of breath, a fever of 38.5°C, and a 7 kg loss of weight within the past month. At the aforementioned hospital, a chest radiograph indicated the presence of bilateral interstitial pneumonia. Subsequent microbiological investigations, including Ziehl-Neelsen staining and a PCR-based assay to test for the presence of M. tuberculosis on expectoration, indicated that there were no signs of such an infection. The patient was then transferred to our department for further evaluation. Clinical examination of the patient verified both a body temperature of 38 – 38.5°C and dyspnoea with 90% oxygen saturation under 6 L/min oxygen. The medical history of the patient was unremarkable, except for previous treatment for arterial hypertension. The total body tomodensitometry indicated the presence of nodules in both lungs, in the mediastinal lymph nodes, and in a right axilar lymph node. The pertinent laboratory assays were performed and indicated a value of 5.9 leucocytes/ml with 76% polymorphonuclear cells and 190 platelets/ml.