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采用单链构象多态性聚合酶链反应（SSCP-PCR）对伊朗霍尔木兹甘省间日疟原虫患者分离株中MSP-1基因的遗传变异进行研究。

Genetic Variation of MSP-1 Gene in Plasmodium vivax Isolated from Patients in Hormozgan Province, Iran using SSCP-PCR.

作者信息

Miahipour A, Keshavarz H, Heidari A, Raeisi A, Rezaeian M, Rezaie S

机构信息

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Parasitol. 2012;7(4):1-7.

PMID:23323085
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3537474/
Abstract

BACKGROUND

The main goal of present study was to detect polymorphism in MSP-1 gene which is a major blood stage candidate for vaccine in Plasmodium vivax by Single Strand Conformational Polymorphism-Polymerase Chain Reaction (SSCP-PCR).

METHODS

During 2008 to 2010 fifty samples were collected from Iranian patients with P. vivax in Hormozgan Province, southern Iran. All of the samples were detected by microscopical examination. Amplification of MSP-1 gene was done by PCR after DNA extraction. Single strand DNAs due to using in SSCP, was electrophoresed on polyacrylamid- Bisacrylamid gel then banding patterns were revealed by silver-staining method. Sequencing as a typing method was performed for some isolates.

RESULTS

All of the 50 isolates were positive microscopically. Totally 12 (24%) isolates showed 440 bp and 38 (76%) showed 500 bp in PCR assay. SSCP analysis revealed four banding patterns. Pattern I (10/50), Pattern II (12/50), Pattern III (27/50), and Pattern IV (1/50). The results sequencing analysis of the MSP-1 gene in 19 isolates revealed diversity in nucleotides and amino acid in Iranian P. vivax isolates.

CONCLUSION

Our study confirms that the SSCP-PCR is a rapid method for detecting polymorphism in MSP-1 gene in P. vivax. The presence of different haplotypes in MSP-1 gene shows that several P. vivax strains exist in malaria endemic areas of Iran.

摘要

背景

本研究的主要目的是通过单链构象多态性-聚合酶链反应（SSCP-PCR）检测间日疟原虫中作为疫苗主要血期候选基因的MSP-1基因的多态性。

方法

2008年至2010年期间，从伊朗南部霍尔木兹甘省的间日疟原虫患者中收集了50份样本。所有样本均通过显微镜检查进行检测。DNA提取后，通过PCR扩增MSP-1基因。由于用于SSCP分析，单链DNA在聚丙烯酰胺-双丙烯酰胺凝胶上进行电泳，然后通过银染法显示条带模式。对一些分离株进行测序作为分型方法。

结果

所有50株分离株显微镜检查均为阳性。在PCR检测中，共有12株（24%）显示440 bp，38株（76%）显示500 bp。SSCP分析显示出四种条带模式。模式I（10/50）、模式II（12/50）、模式III（27/50）和模式IV（1/50）。对19株分离株的MSP-1基因进行测序分析的结果显示，伊朗间日疟原虫分离株在核苷酸和氨基酸方面存在多样性。

结论

我们的研究证实，SSCP-PCR是检测间日疟原虫MSP-1基因多态性的快速方法。MSP-1基因中不同单倍型的存在表明伊朗疟疾流行地区存在几种间日疟原虫菌株。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/1281dfe2dec5/IJPA-7-001-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/da6e5cd4bbf5/IJPA-7-001-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/0219d079c731/IJPA-7-001-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/ee61eb811e22/IJPA-7-001-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/1281dfe2dec5/IJPA-7-001-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/da6e5cd4bbf5/IJPA-7-001-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/0219d079c731/IJPA-7-001-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/ee61eb811e22/IJPA-7-001-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5a0/3537474/1281dfe2dec5/IJPA-7-001-g004.jpg

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