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Diagnosis of cutaneous tuberculosis in biopsy specimens by PCR and southern blotting.

机译:PCR和Southern blotting对活检标本中皮肤结核的诊断。

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摘要

AIMS: To evaluate the use of a gene amplification and hybridisation method for detecting mycobacterial nucleic acid as a possible diagnostic method for cutaneous tuberculosis infection. METHODS: Biopsy specimens from 20 patients with various skin conditions of possible tuberculous aetiology were studied. Six patients had ulcerative nodules, seven lupiform lesions, two non-necrotic granulomas, one scrofulous lichen, one impetigo, one erythematosus lesions, one warty lesions, and one suspected tuberculous lipoma. Biopsy specimens were stained using Ziehl-Neelsen stain and cultured in Lowenstein-Jensen medium. DNA was extracted and then amplified by PCR using primers specific for the Mycobacterium tuberculosis complex. Specificity was confirmed by Southern blotting. RESULTS: Of the specimens, 30% were positive for mycobacteria on staining with Ziehl-Neelsen stain, 60% were culture positive and 85% PCR positive. Only 35.2% of specimens were positive with all three techniques. A further 32.5% were both culture and PCR positive. All PCR negative samples were also negative when cultured or stained with Ziehl-Neelsen stain. Of the PCR positive specimens, 29.4% were negative when cultured or stained. CONCLUSIONS: PCR, using suitable primers, is an efficient and sensitive method for the diagnosis of cutaneous tuberculosis.
机译:目的:评估基因扩增和杂交方法用于检测分枝杆菌核酸作为皮肤结核感染的可能诊断方法的用途。方法:对20例皮肤病患者的活检标本进行了研究,这些患者可能患有结核病。 6例患者有溃疡性结节,7例肾形病变,2例非坏死性肉芽肿,1例阴囊性苔藓,1例脓疱病,1例红斑病,1例疣性病变和1例疑似结核性脂瘤。活检标本用Ziehl-Neelsen染色剂染色,并在Lowenstein-Jensen培养基中培养。提取DNA,然后使用对结核分枝杆菌复合物具有特异性的引物通过PCR进行扩增。通过Southern印迹证实了特异性。结果:标本中,用Ziehl-Neelsen染色染色的分枝杆菌阳性率为30%,培养阳性,PCR阳性率为85%。三种技术中只有35.2%的标本呈阳性。另外32.5%的培养物和PCR均为阳性。培养或用Ziehl-Neelsen染色后,所有PCR阴性样品也均为阴性。在PCR阳性样本中,培养或染色后阴性的样本为29.4%。结论:使用合适的引物进行PCR是诊断皮肤结核的有效且灵敏的方法。

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