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印度拉贾斯坦邦斋浦尔市两个贫民窟分离株的遗传多样性和药敏谱

Genetic diversity & drug sensitivity profiles of isolates from two slums of Jaipur city, Rajasthan, India.

作者信息

Malhotra Bharti, Dashora Deepti, Kumar Vipin, Goyal Sumit, Sharma Bhavana, Kumar Madhu, Gupta Kailash Narayan, Sharma Vishnu Dutt, Chauhan D S, Katoch Kiran, Katoch Vishwa Mohan

机构信息

Department of Microbiology & Immunology, SMS Medical College, Rajasthan, India.

Department of Microbiology & Immunology, SMS Medical College; Department of Zoology, Cell & Molecular Biology Laboratory, University of Rajasthan, Rajasthan, India.

出版信息

Indian J Med Res. 2017 Jan;145(1):74-83. doi: 10.4103/ijmr.IJMR_336_14.

Abstract

BACKGROUND & OBJECTIVES: Slums are considered as hotspots of tuberculosis (TB). The study of genetic diversity and drug susceptibility profile of Mycobacterium tuberculosis (MTB) will help understand the transmission dynamics and can be used for better prevention and control of the disease. The aim of this study was to determine the drug susceptibility profiles and genetic diversity using the random amplified polymorphic DNA (RAPD) and mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU VNTR) of MTB isolates from sputum samples of pulmonary TB patients residing in the two slums of Jaipur city in Rajasthan, India.

METHODS

Sputum samples collected from pulmonary TB patients, their contacts and suspects during 2010-2012 were processed for microscopy and mycobacterial culture. Drug susceptibility testing was done by one per cent indirect proportion method on Lowenstein-Jensen medium for first-line anti-TB drugs rifampicin, isoniazid, ethambutol and streptomycin. MTB DNA was extracted by physicochemical method, and DNA fingerprinting was done by RAPD and MIRU VNTR analysis.

RESULTS

Among 175 sputum samples collected, 75 were positive (43.8%) for acid-fast bacilli, 83 for MTB culture and four were contaminated. Fifty two isolates (62.7%) were fully sensitive to four drugs, and five (6%) were multidrug resistant (MDR). RAPD analysis of 81 isolates revealed six clusters containing 23 (28.4%) isolates, and 58 (71.6%) were unique. MIRU VNTR analysis clustered 20 (24.7%) isolates, and 61 (75.3%) were unique.

INTERPRETATION & CONCLUSIONS: About 62.7 per cent isolates from the sputum samples from slum areas were sensitive to four drugs; six per cent of isolates were MDR. Poly-resistance other than MDR was high (16%). About one-fourth isolates were clustered by either method. RAPD was rapid, less expensive but had low reproducibility. MIRU VNTR analysis could identify to greater extent the epidemiological link in the population studied.

摘要

背景与目的

贫民窟被视为结核病(TB)的高发区。对结核分枝杆菌(MTB)的遗传多样性和药敏谱进行研究,将有助于了解其传播动态,并可用于更好地预防和控制该疾病。本研究的目的是利用随机扩增多态性DNA(RAPD)和分枝杆菌散布重复单位 - 可变数目串联重复序列(MIRU VNTR),对来自印度拉贾斯坦邦斋浦尔市两个贫民窟的肺结核患者痰标本中的MTB分离株进行药敏谱和遗传多样性分析。

方法

收集2010 - 2012年期间肺结核患者、其接触者及疑似患者的痰标本,进行显微镜检查和分枝杆菌培养。采用1%间接比例法在罗 - 琴培养基上对一线抗结核药物利福平、异烟肼、乙胺丁醇和链霉素进行药敏试验。通过物理化学方法提取MTB DNA,并采用RAPD和MIRU VNTR分析进行DNA指纹图谱分析。

结果

在收集的175份痰标本中,75份(43.8%)抗酸杆菌阳性,83份MTB培养阳性,4份污染。52株分离株(62.7%)对四种药物完全敏感,5株(6%)为耐多药(MDR)。对81株分离株进行RAPD分析,发现6个聚类,包含23株(28.4%)分离株,58株(71.6%)为独特型。MIRU VNTR分析将20株(24.7%)分离株聚类,61株(75.3%)为独特型。

解读与结论

来自贫民窟地区痰标本的分离株中,约62.7%对四种药物敏感;6%的分离株为MDR。除MDR外的多耐药情况较高(16%)。约四分之一的分离株可通过两种方法中的任何一种聚类。RAPD快速、成本较低,但重复性差。MIRU VNTR分析在更大程度上能够识别所研究人群中的流行病学联系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4af6/5460578/fb61324822bd/IJMR-145-74-g003.jpg

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