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通过靶向RLEP序列的聚合酶链反应从土壤样本中检测麻风分枝杆菌DNA。

Detection of Mycobacterium leprae DNA from soil samples by PCR targeting RLEP sequences.

作者信息

Lavania Mallika, Katoch K, Sachan Pawan, Dubey Anuraag, Kapoor Shashi, Kashyap Meera, Chauhan D S, Singh H B, Sharma V D, Jadhav R S, Katoch V M

机构信息

National JALMA Institute for Leprosy & Other Mycobacterial Diseases (ICMR), Taj Ganj, Agra 282001.

出版信息

J Commun Dis. 2006 Mar;38(3):269-73.

Abstract

Despite near elimination of leprosy as a public health problem, several problems in leprosy still remain. These include early detection, determining efficacy of the treatment and differentiating relapses from re-infection. These aspects have important impact on the patients undergoing treatment and also have a bearing on understanding transmission dynamics in the community. While early diagnosis and management do not need major technological inputs, various reports have suggested that M. leprae is found in the environment and may have a role in continued transmission of disease. In earlier studies from other parts of world the presence of M. leprae DNA in the environment has been investigated both by microbiological and molecular studies. In the present study, an attempt was made to extract M. leprae DNA from soil samples, which were collected from eighteen different locations including 3 from our Institute area and 15 from different villages of Ghatampur area. We optimized a protocol for the extraction of DNA and amplified a fragment of M. leprae using specific primers targeting RLEP sequences. It was found that 33.3% of these soil samples collected from areas inhabited by leprosy cases gave positive result for M. leprae specific DNA. The utility of this method needs to be explored on a larger scale to establish the presence of M.leprae in the environment, and its role in the spread of the disease.

摘要

尽管麻风病作为一个公共卫生问题已近乎消除,但麻风病仍存在若干问题。这些问题包括早期检测、确定治疗效果以及区分复发与再次感染。这些方面对正在接受治疗的患者有重要影响,也关系到对社区传播动态的理解。虽然早期诊断和管理不需要重大技术投入,但各种报告表明麻风杆菌存在于环境中,可能在疾病的持续传播中起作用。在世界其他地区的早期研究中,已通过微生物学和分子研究对环境中麻风杆菌DNA的存在情况进行了调查。在本研究中,我们尝试从土壤样本中提取麻风杆菌DNA,这些土壤样本采自18个不同地点,其中3个来自我们研究所区域,15个来自加坦布尔地区的不同村庄。我们优化了DNA提取方案,并使用靶向RLEP序列的特异性引物扩增了麻风杆菌的一个片段。结果发现,从麻风病患者居住地区采集的这些土壤样本中,33.3%的样本对麻风杆菌特异性DNA检测呈阳性。需要更大规模地探索该方法的实用性,以确定环境中麻风杆菌的存在情况及其在疾病传播中的作用。

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