Cruz Andrezza Furquim da, Furini Renata Bazan, Roselino Ana Maria Ferreira
Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Brasil.
An Bras Dermatol. 2011 Jul-Aug;86(4):651-6. doi: 10.1590/s0365-05962011000400004.
The Polymerase Chain Reaction (PCR) technique has been frequently used in the molecular diagnosis of leprosy.
To compare the results of PCR with four pairs of Mycobacterium leprae specific primers as well as to compare these results to multibacillary (MB) and paucibacillary (PB) leprosy according to the WHO operational classification.
28 DNA samples, collected from the frozen skin biopsies and biopsy imprints on filter paper of 23 patients (14 MB and PB 9), were examined for PCR using primers which amplify 131, 151 and 168bp of specific microsatellite regions and a 336 fragment of the Ml MntH (ML2098) gene.
M.leprae bacillus could be detected in 22 (78.6%) of the 28 samples. 9 (45%) of the 20 biopsy samples and 6 (75%) of the 8 imprints were positive to TTC. 7 (35.5%) skin biopsy specimens and 5 (62.5%) imprints were positive to AGT, and 11 (55%) biopsies and 4 (50%) were positive to AGT. 11 (55%) skin biopsies and 4 (50%) imprints were positive to AT. 8(38%) skin biopsies and 5 (62.5%) imprints were positive to the Ml MntH gene. In the MB group, the microsatellites detected the bacillus in 78.5% of the samples, and the Ml MntH gene in 57.1% of the samples, independent of the clinical material. In the PB group 55.5% of samples were positive to the microsatellite primers, while 22.2% were positive to the Ml MntH gene.
These results show that both the specific regions of microsatellites, as well as the Ml MntH gene fragment can be useful tools for detecting the M. leprae DNA by PCR in frozen skin biopsy samples and filter paper biopsy imprints.
聚合酶链反应（PCR）技术已频繁用于麻风病的分子诊断。
比较使用四对麻风分枝杆菌特异性引物进行PCR的结果，并根据世界卫生组织的操作分类将这些结果与多菌型（MB）和少菌型（PB）麻风病进行比较。
从23例患者（14例MB和9例PB）的冷冻皮肤活检标本和滤纸上的活检印记中收集28份DNA样本，使用能扩增特定微卫星区域131、151和168bp以及Ml MntH（ML2098）基因336片段的引物进行PCR检测。
28份样本中有22份（78.6%）检测到麻风杆菌。20份活检样本中有9份（45%）和8份印记中有6份（75%）对TTC呈阳性。7份（35.5%）皮肤活检标本和5份（62.5%）印记对AGT呈阳性，11份（55%）活检样本和4份（50%）对AGT呈阳性。11份（55%）皮肤活检样本和4份（50%）印记对AT呈阳性。8份（38%）皮肤活检样本和5份（62.5%）印记对Ml MntH基因呈阳性。在MB组中，微卫星在78.5%的样本中检测到杆菌，Ml MntH基因在57.1%的样本中检测到，与临床材料无关。在PB组中，55.5%的样本对微卫星引物呈阳性，而22.2%的样本对Ml MntH基因呈阳性。
这些结果表明，微卫星特定区域以及Ml MntH基因片段均可作为通过PCR检测冷冻皮肤活检样本和滤纸上活检印记中麻风杆菌DNA的有用工具。
