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使用插入序列6110(IS6110)和电泳中Rm 0.64的卡介苗分枝杆菌蛋白组分进行多重聚合酶链反应,用于诊断结核性淋巴结炎的靶基因。

Multiplex polymerase chain reaction using insertion sequence 6110 (IS6110) and mycobacterial protein fraction from BCG of Rm 0.64 in electrophoresis target genes for diagnosis of tuberculous lymphadenitis.

作者信息

Sharma K, Gupta N, Sharma A, Singh G, Gupta P K, Rajwanshi A, Varma S C, Sharma M

机构信息

Department of Medical Microbiology, PGIMER, Chandigarh, India.

出版信息

Indian J Med Microbiol. 2013 Jan-Mar;31(1):24-8. doi: 10.4103/0255-0857.108714.

Abstract

PURPOSE

Tubercular lymphadenitis (TBLA) is a common manifestations of extrapulmonary tuberculosis (EPTB) accounting for 30-40% of cases. Prompt diagnosis and timely initiation of anti-tubercular therapy (ATT) is the key for successful clinical outcome. This study was carried out to evaluate multiplex polymerase chain reaction (MPCR) using MPB64 and IS6110, and compare with the conventional methods for rapid diagnosis of TBLA.

MATERIALS AND METHODS

In our study, lymph node fine-needle aspirates of patients were evaluated for TBLA. They were classified as Group I: TBLA group, divided into (a) Confirmed TBLA cases (n0 = 80): Culture/smear-positive or cytological examination showing presence of epithelioid cell granuloma with or without multinucleate giant cell and caseation necrosis with presence of AFB, and (b) suspected TBLA cases ( n = 30): Culture/smear-negative and cytological examination showing presence of epithelioid cell granuloma and response to ATT and Group II (Control) (n = 25): Patients of lymphadenopathy confirmed to be caused by other diseases such as sarcoidosis, lymphoma, etc., All samples were subjected to conventional tests and MPCR. For MPCR we used Mycobacterium tuberculosis-specific deoxyribonucleic acid sequences specific for the MPB64 and IS6110 region.

RESULTS

In the confirmed TBLA group, Ziehl-Neelsen (ZN) smear, cytology, culture, and MPCR positivity was 30%, 70%, 26.3%, and 91.3% respectively. In the suspected TBLA group, smear and culture were negative, and sensitivity of cytology and MPCR was 73.3% and 86.6%, respectively. In the control group all tests were found to be negative, thus giving a specificity of 100% to all the tests in the study.

CONCLUSION

In conclusion, techniques like MPCR with high sensitivity and specificity can play an important role in rapid diagnosis of TBLA.

摘要

目的

结核性淋巴结炎(TBLA)是肺外结核病(EPTB)的常见表现,占病例的30%-40%。及时诊断并及时开始抗结核治疗(ATT)是取得成功临床结果的关键。本研究旨在评估使用MPB64和IS6110的多重聚合酶链反应(MPCR),并与传统方法比较以快速诊断TBLA。

材料与方法

在我们的研究中,对患者的淋巴结细针穿刺抽吸物进行TBLA评估。它们被分为I组:TBLA组,分为(a)确诊TBLA病例(n0 = 80):培养/涂片阳性或细胞学检查显示存在上皮样细胞肉芽肿,伴有或不伴有多核巨细胞以及存在抗酸杆菌的干酪样坏死,以及(b)疑似TBLA病例(n = 30):培养/涂片阴性且细胞学检查显示存在上皮样细胞肉芽肿以及对ATT有反应;II组(对照组)(n = 25):经证实由结节病、淋巴瘤等其他疾病引起的淋巴结病患者。所有样本均进行传统检测和MPCR。对于MPCR,我们使用针对MPB64和IS6110区域的结核分枝杆菌特异性脱氧核糖核酸序列。

结果

在确诊TBLA组中,萋-尼(ZN)涂片、细胞学、培养和MPCR阳性率分别为30%、70%、26.3%和91.3%。在疑似TBLA组中,涂片和培养均为阴性,细胞学和MPCR的敏感性分别为73.3%和86.6%。在对照组中,所有检测均为阴性,因此本研究中所有检测的特异性均为100%。

结论

总之,像MPCR这样具有高敏感性和特异性的技术在TBLA的快速诊断中可发挥重要作用。

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