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通过聚合酶链反应(PCR)和Southern印迹法对活检标本中的皮肤结核进行诊断。

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

作者信息

Quirós E, Maroto M C, Bettinardi A, González I, Piédrola G

机构信息

Department of Microbiology, Medical School, University of Granada, Spain.

出版信息

J Clin Pathol. 1996 Nov;49(11):889-91. doi: 10.1136/jcp.49.11.889.

Abstract

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.

摘要

目的

评估一种基因扩增和杂交方法检测分枝杆菌核酸作为皮肤结核感染可能诊断方法的应用。

方法

对20例有各种可能结核病因皮肤状况的患者的活检标本进行研究。6例有溃疡性结节,7例有狼疮样损害,2例有非坏死性肉芽肿,1例有瘰疬性苔藓,1例有脓疱疮,1例有红斑性损害,1例有疣状损害,1例有疑似结核性脂肪瘤。活检标本用萋-尼染色并在罗-琴培养基中培养。提取DNA,然后使用针对结核分枝杆菌复合群的引物通过聚合酶链反应(PCR)进行扩增。通过Southern印迹法确认特异性。

结果

在这些标本中,萋-尼染色分枝杆菌阳性的占30%,培养阳性的占60%,PCR阳性的占85%。仅35.2%的标本三种技术均为阳性。另外32.5%的标本培养和PCR均为阳性。所有PCR阴性的样本在培养或萋-尼染色时也为阴性。在PCR阳性的标本中,29.4%在培养或染色时为阴性。

结论

使用合适的引物,PCR是诊断皮肤结核的一种有效且敏感的方法。

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