Differentiation of BCG-induced lymphadenitis from tuberculosis in lymph node biopsy specimens by molecular analyses ofpncA andoxyR
Department of Pathology, National Cheng Kung University Hospital, Tainan, Taiwan.The Journal of Pathology (Impact Factor: 7.43). 01/1998; 184(1):96-102. DOI: 10.1002/(SICI)1096-9896(199801)184:1<96::AID-PATH989>3.0.CO;2-M
Without culture, differentiation of bacille Calmette-Guérin-induced lymphadenitis (BCG-LA) from tuberculosis (TB) is sometimes difficult by histology, but is important because of different treatment schemes. The purpose of this study was to investigate the feasibility of differentiating BCG-LA from TB in lymph nodes (LNs) by molecular analyses of two recently identified genes, pncA and oxyR. In both genes, a single tuberculosis difference exists between Mycobacterium bovis and M. tuberculosis. M tuberculosis complex (MTC) DNA was first detected in nine of ten formalin-fixed, paraffin-embedded LNs from patients aged under 20 years with suspected mycobacterial infections, using polymerase chain reaction (PCR) for IS6110, an insertion sequence specific for MTC species. PCR, together with direct DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) assay, was then performed to identify polymorphic nucleotide in pncA and oxyR, respectively. For comparison, 37 adult cases of tuberculous lymphadenitis were also analysed by PCR-single strand conformation polymorphism (SSCP) assay for pncA and by PCR-RFLP for oxyR. The results revealed that five of the nine IS6110-positive child cases had a G residue at nucleotide 169 in pncA, and also had a three-band pattern after digesting the amplified oxyR segment with AluI, suggesting BCG-LA. The remaining four child cases, as well as all adult cases with detectable IS6110, showed no motility shift in pncA PCR-SSCP and had the same one-band pattern as M. tuberculosis in oxyR PCR-RFLP, suggesting TB lymphadenitis. The data from molecular analyses showed a good correlation with the vaccination history and clinicopathological findings, except for one case. This study indicates that molecular assay of either oxyR or pncA could be a rapid and useful tool to distinguish BCG-LA from TB.
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ABSTRACT: In its role in the presentation of foreign antigens to the immune system, the lymph node is frequently involved in both local and systemic infections. This may manifest as lymphadenopathy with reactive changes or localized infectious lymphadenitis, with or without necrosis and granulomatous inflammation, depending on the infectious agent. The vast majority of these infections are routinely diagnosed using nonmolecular methods, including culture, serology, and antigen testing. However, in some cases, other methods may be unavailable, or the presentation and appearance may be atypical, and direct confirmation of the infecting agent by in situ hybridization (ISH) or amplified nucleic acid detection is desired in the lymph node tissue itself.
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ABSTRACT: Mycobacterium genavense is a recently described fastidious mycobacterium identified as a pathogen causing disseminated infection in patients with advanced human immunodeficiency virus (HIV) disease. In this report, we describe the first reported case of disseminated M. genavense infection in a patient with acquired immunodeficiency syndrome (AIDS) in Taiwan. A 22-year-old Chinese man was found to be seropositive for HIV at age 18, in 1993. In 1997, he presented with abdominal pain, weight loss, low CD4 lymphocyte count, hepatomegaly, and generalized lymphadenopathy. Microscopic examination of a biopsy specimen from an inguinal lymph node showed both ill- and well-formed noncaseating granulomas. Numerous acid-fast bacilli were present in the histiocyte cytoplasm. Although the organism did not grow on conventional solid media used in our laboratory, two molecular biology techniques, including polymerase chain reaction (PCR) followed by sequencing of 16S rRNA, and PCR together with restriction enzyme fragment polymorphism analysis, confirmed the M. genavense infection. The patient's abdominal symptoms responded well to a chemotherapy regimen that included ethambutol, ciprofloxacin, and clarithromycin, and he survived more than 6 months after diagnosis. However, the lymphadenopathy was still present at his final follow-up. Our report indicates that disseminated infection with M. genavense should be added to the list of differential diagnoses of secondary infections in advanced AIDS patients in Taiwan.Journal of the Formosan Medical Association 02/1999; 98(1):62-5. · 1.97 Impact Factor
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