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基于聚合酶链反应检测巴基斯坦临床幽门螺杆菌菌株中的相变基因

PCR Based Detection of Phase Variable Genes in Pakistani Based Clinical Helicobacter pylori Strains.

作者信息

Ahmad Sajjad, Ahmad Faisal, Rahman Faiz Ur, Khan Salman, Murad Waheed, Mughal Imran, Ur Rahman Amjad, Muhammad Khan Fida, Khan Imad, Ahmad Hajra

机构信息

Department of Microbiology, Faculty of Biological Sciences, Quaid-i-azam University, Islamabad, Pakistan.

Department of Microbiology and Molecular Genetics (MMG), University of Punjab, Lahore, Pakistan.

出版信息

Jundishapur J Microbiol. 2016 Jul 3;9(7):e31824. doi: 10.5812/jjm.31824. eCollection 2016 Jul.

DOI:10.5812/jjm.31824
PMID:27679701
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5035388/
Abstract

BACKGROUND

The distribution pattern of phase-variable genes varies from strain to strain and from region to region. The present study was carried out to investigate the distribution pattern of phase-variable genes within Pakistan-based Helicobacter pylori strains and to analyze and compare them with strains prevalent in other parts of the world.

OBJECTIVES

To determine the distribution pattern of phase-variable genes in H. pylori strains circulating in Pakistan.

PATIENTS AND METHODS

Biopsy samples were collected from 85 symptomatic patients suffering from various upper gastrointestinal tract symptoms. The biopsy specimens were chopped, then inoculated on H. pylori-specific media and incubated in a Campylobacter Gas Generating kit. Positive isolates were further confirmed via staining and biochemical procedures. Primers were designed for five phase-variable genes using OligoCalc, an oligonucleotide properties calculator (version 3.26) according to parameters stipulated in the literature. Polymerase chain reaction (PCR) was performed on all positive isolates to determine the presence or absence of phase-variable genes.

RESULTS

On culturing, the prevalence of H. pylori infections in the samples was 44.7%. The prevalence was higher in females than in males, and it increased with age. PCR amplification revealed that the hsdR gene was present in 79% of samples, while the mod and β-subunit genes were present in 16% and 30% of samples, respectively. The streptococcal M protein gene was found in 79%, while the fliP gene was prevalent in 56%.

CONCLUSIONS

The distribution patterns of phase-variable genes in Pakistani H. pylori strains were found to be somewhat different. The dominant prevalence of the hsdR gene was an interesting finding, considering its role in bacterial defense in both micro- and macroenvironments.

摘要

背景

相位可变基因的分布模式因菌株和地区而异。本研究旨在调查巴基斯坦幽门螺杆菌菌株中相位可变基因的分布模式，并与世界其他地区流行的菌株进行分析和比较。

目的

确定在巴基斯坦流行的幽门螺杆菌菌株中相位可变基因的分布模式。

患者与方法

从85名患有各种上消化道症状的有症状患者中采集活检样本。将活检标本切碎，然后接种在幽门螺杆菌特异性培养基上，并在弯曲杆菌气体发生试剂盒中孵育。通过染色和生化程序进一步确认阳性分离株。根据文献规定的参数，使用寡核苷酸特性计算器OligoCalc（版本3.26）为五个相位可变基因设计引物。对所有阳性分离株进行聚合酶链反应（PCR），以确定相位可变基因的存在与否。

结果

培养后，样本中幽门螺杆菌感染的患病率为44.7%。女性患病率高于男性，且随年龄增长而增加。PCR扩增显示，79%的样本中存在hsdR基因，而mod和β亚基基因分别存在于16%和30%的样本中。79%的样本中发现了链球菌M蛋白基因，而fliP基因在56%的样本中普遍存在。

结论

发现巴基斯坦幽门螺杆菌菌株中相位可变基因的分布模式有所不同。考虑到hsdR基因在微观和宏观环境中细菌防御中的作用，其占主导地位的患病率是一个有趣的发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/60e02059328e/jjm-09-07-31824-i003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/703d3ab49c80/jjm-09-07-31824-i001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/4c1e1fa0c90a/jjm-09-07-31824-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/55c8cff58bf1/jjm-09-07-31824-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/64ce88265d4f/jjm-09-07-31824-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/b0614494f8c9/jjm-09-07-31824-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/f1048de5800a/jjm-09-07-31824-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/60e02059328e/jjm-09-07-31824-i003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/703d3ab49c80/jjm-09-07-31824-i001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/7c38c3ec3bfd/jjm-09-07-31824-i002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/4c1e1fa0c90a/jjm-09-07-31824-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/55c8cff58bf1/jjm-09-07-31824-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/64ce88265d4f/jjm-09-07-31824-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/b0614494f8c9/jjm-09-07-31824-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/f1048de5800a/jjm-09-07-31824-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae4f/5035388/60e02059328e/jjm-09-07-31824-i003.jpg

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