02774nas a2200349 4500000000100000008004100001260001300042653002800055653001600083653001900099653001400118653001700132653001400149653001100163653001800174653002500192653003100217653003200248653002300280653003000303653001700333100001200350700001300362700001200375700001400387700001000401245016800411300001000579490000700589520181400596022001402410 1997 d c1997 Jan10aDiagnosis, Differential10aDNA Primers10aDNA, Bacterial10aFixatives10aFormaldehyde10aGranuloma10aHumans10aMycobacterium10aMycobacterium leprae10aMycobacterium tuberculosis10aNontuberculous Mycobacteria10aParaffin Embedding10apolymerase chain reaction10aTuberculosis1 aOsaki M1 aAdachi H1 aGomyo Y1 aYoshida H1 aIto H00aDetection of mycobacterial DNA in formalin-fixed, paraffin-embedded tissue specimens by duplex polymerase chain reaction: application to histopathologic diagnosis. a78-830 v103 a

Granuloma is a chronic inflammatory process associated with noninfectious agents or infectious diseases such as tuberculosis. Determination of the causative agent might be occasionally difficult in histopathologic sections. In this study, we examined 60 specimens of granuloma or inflammatory lesions that were originally diagnosed as 51 cases of granulomatous inflammation, 6 of leprosy, and 3 of atypical mycobacteriosis. The diagnoses in the last two categories were made both histologically and clinically. All of the sections and DNA were prepared from formalin-fixed, paraffin-embedded blocks. Histopathologic and immunohistochemical findings were compared with the results of duplex polymerase chain reaction (PCR) using two primers to amplify mycobacterial-common 383-base pair (bp) DNA and Mycobacterium tuberculosis-complex-specific 240-bp DNA. Six samples of leprosy and three of atypical mycobacteriosis showed the 383-bp but not the 240-bp band. Among the 51 specimens of granulomatous inflammations, nine showed no band of even the beta-globin, the cases being excluded from this analysis. The 42 specimens of granulomatous inflammation were subdivided into three categories by PCR: (1) 383- and 240-bp positive; (2) 383-bp positive and 240-bp negative; and (3) both negative. Category 1 included 32 specimens (76.2%), being considered as tuberculosis. One specimen was classified into Category 2, indicating possible atypical mycobacterium. Category 3 included nine specimens, composed of five of sarcoidosis and four other agent-induced granulomas, when compared with histologic and clinical findings. These findings indicate that the PCR assay using DNA extracted from paraffin-embedded materials provides useful information to differentiate tuberculosis from other type of granulomas.

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