Joshi Hema, Valecha Neena, Verma Anju, Kaul Asha, Mallick Prashant K, Shalini Sneh, Prajapati Surendra K, Sharma Surya K, Dev Vas, Biswas Sukla, Nanda Nutan, Malhotra M S, Subbarao Sarala K, Dash Aditya P
National Institute of Malaria Research (ICMR), 22-Sham Nath Marg, Delhi, India.
Malar J. 2007 May 21;6:60. doi: 10.1186/1475-2875-6-60.
Molecular techniques have facilitated the studies on genetic diversity of Plasmodium species particularly from field isolates collected directly from patients. The msp-1 and msp-2 are highly polymorphic markers and the large allelic polymorphism has been reported in the block 2 of the msp-1 gene and the central repetitive domain (block3) of the msp-2 gene. Families differing in nucleotide sequences and in number of repetitive sequences (length variation) were used for genotyping purposes. As limited reports are available on the genetic diversity existing among Plasmodium falciparum population of India, this report evaluates the extent of genetic diversity in the field isolates of P. falciparum in eastern and north-eastern regions of India.
A study was designed to assess the diversity of msp-1 and msp-2 among the field isolates from India using allele specific nested PCR assays and sequence analysis. Field isolates were collected from five sites distributed in three states namely, Assam, West Bengal and Orissa.
P. falciparum isolates of the study sites are highly diverse in respect of length as well as sequence motifs with prevalence of all the reported allelic families of msp-1 and msp-2. Prevalence of identical allelic composition as well as high level of sequence identity of alleles suggest a considerable amount of gene flow between the P. falciparum populations of different states. A comparatively higher proportion of multiclonal isolates as well as multiplicity of infection (MOI) was observed among isolates of highly malarious districts Karbi Anglong (Assam) and Sundergarh (Orissa). In all the five sites, R033 family of msp-1 was observed to be monomorphic with an allele size of 150/160 bp. The observed 80-90% sequence identity of Indian isolates with data of other regions suggests that Indian P. falciparum population is a mixture of different strains.
The present study shows that the field isolates of eastern and north-eastern regions of India are highly diverse in respect of msp-1 (block 2) and msp-2 (central repeat region, block 3). As expected Indian isolates present a picture of diversity closer to southeast Asia, Papua New Guinea and Latin American countries, regions with low to meso-endemicity of malaria in comparison to African regions of hyper- to holo-endemicity.
分子技术推动了疟原虫物种遗传多样性的研究,特别是对直接从患者采集的野外分离株的研究。msp-1和msp-2是高度多态性标记,msp-1基因的第2区和msp-2基因的中央重复区(第3区)已报道有大量等位基因多态性。核苷酸序列和重复序列数量(长度变异)不同的家族被用于基因分型。由于关于印度恶性疟原虫群体中存在的遗传多样性的报道有限,本报告评估了印度东部和东北部地区恶性疟原虫野外分离株的遗传多样性程度。
设计一项研究,使用等位基因特异性巢式PCR分析和序列分析来评估印度野外分离株中msp-1和msp-2的多样性。野外分离株从分布在阿萨姆邦、西孟加拉邦和奥里萨邦三个邦的五个地点采集。
研究地点的恶性疟原虫分离株在长度以及序列基序方面高度多样,msp-1和msp-2的所有已报道等位基因家族均有流行。相同等位基因组成的流行以及等位基因的高水平序列同一性表明不同邦的恶性疟原虫群体之间存在相当数量的基因流动。在疟疾高发地区卡尔比安隆(阿萨姆邦)和孙德尔加尔(奥里萨邦)的分离株中观察到相对较高比例的多克隆分离株以及感染复数(MOI)。在所有五个地点,观察到msp-1的R033家族是单态的,等位基因大小为150/160 bp。印度分离株与其他地区数据的序列同一性为80 - 90%,这表明印度恶性疟原虫群体是不同菌株的混合体。
本研究表明,印度东部和东北部地区的野外分离株在msp-1(第2区)和msp-2(中央重复区,第3区)方面高度多样。正如预期的那样,与非洲高流行至全流行地区相比,印度分离株呈现出更接近东南亚、巴布亚新几内亚和拉丁美洲国家(疟疾低至中度流行地区)的多样性情况。