發(fā)布時(shí)間：2018-04-22 10:15

  本文選題：惡性瘧原蟲(chóng) + 微衛(wèi)星��； 參考：《中國(guó)寄生蟲(chóng)學(xué)與寄生蟲(chóng)病雜志》2017年03期

【摘要】：目的研究中緬邊境拉咱地區(qū)和泰國(guó)東南Srisaket、Trat和Ranong等3個(gè)地區(qū)惡性瘧原蟲(chóng)(Plasmodium falciparum)K13基因側(cè)翼微衛(wèi)星的多態(tài)性。方法取前期經(jīng)基因型研究的惡性瘧原蟲(chóng)樣品42份,其中中緬邊境拉咱地區(qū)22份(K13基因R539T突變型和野生型各11份),泰國(guó)3個(gè)地區(qū)20份(K13基因R539T突變型9份和野生型11份),對(duì)上述樣品K13基因上游31 900、6 360、150 bp和下游1 700、11 000、31 500 bp共6個(gè)位點(diǎn)的微衛(wèi)星片段進(jìn)行PCR擴(kuò)增,PCR產(chǎn)物經(jīng)毛細(xì)管電泳儀檢測(cè)片段長(zhǎng)度,并采用Bio Calculator軟件進(jìn)行數(shù)據(jù)分析,Gen ALEx軟件計(jì)算等位基因頻率和期望雜合度(He)。結(jié)果中緬邊境拉咱地區(qū)和泰國(guó)3個(gè)地區(qū)惡性瘧原蟲(chóng)蟲(chóng)株K13基因兩側(cè)的6個(gè)微衛(wèi)星位點(diǎn)共檢測(cè)到46個(gè)等位基因,在上游6 360 bp位點(diǎn)的等位基因長(zhǎng)度分別集中在286和278 bp,等位基因頻率為73%和67%。上游31 900 bp位點(diǎn)等位基因長(zhǎng)度分別集中在211和205 bp,等位基因頻率為55%和44%。上游150 bp及下游1 700 bp位點(diǎn)等位基因長(zhǎng)度主要集中在195和209 bp,其他兩個(gè)位點(diǎn)未出現(xiàn)集中分布的情況。中緬邊境拉咱地區(qū)蟲(chóng)株的單倍體型為H1至H4型,泰國(guó)3個(gè)地區(qū)蟲(chóng)株除了2株H2單體型外,其余均為H5和H6型。42份樣品的6個(gè)微衛(wèi)星位點(diǎn)的平均He分別為0.49±0.23、0.42±0.33和0.72±0.12、0.67±0.11。R539T突變的蟲(chóng)株在毗鄰K13基因的微衛(wèi)星位點(diǎn)處He明顯降低,在上游150 bp處分別為0.17和0,在下游1 700 bp處分別為0.3和0。結(jié)論中緬邊境拉咱地區(qū)及泰國(guó)3個(gè)地區(qū)K13基因?yàn)镽539T突變型的惡性瘧原蟲(chóng)蟲(chóng)株均受到青蒿素選擇壓力,等位基因型在上游6 360和31 900 bp兩個(gè)位點(diǎn)上具有明顯的差異。
[Abstract]:Objective to study the polymorphism of Plasmodium falciparum)K13 gene flanking microsatellite in Laasan region of Sino-Myanmar border and Srisakettrat and Ranong in southeast Thailand. Methods 42 samples of Plasmodium falciparum were collected from genotypic study. There were 22 R539T mutant and 11 wild type in R539T gene, 9 in R539T mutation and 11 in wild type in three regions of Thailand. The upstream of K13 gene was 31,900-6 360150 BP and the downstream of K13 gene was 1,7000.11. The fragment length was detected by capillary electrophoresis after PCR amplification of the microsatellite fragment with a total of 6 loci (31,500 BP), and the length of the fragment was determined by capillary electrophoresis. The allele frequency and expected heterozygosity were calculated by Bio Calculator software. Results A total of 46 alleles were detected at 6 microsatellite loci on both sides of the K13 gene of Plasmodium falciparum strain in Laasan region of the Sino-Myanmar border and 3 regions in Thailand. The allele length of the 6360 BP locus in the upstream was 286 and 278 BP, respectively. The allele frequencies were 73% and 67%, respectively. The alleles at 31,900 BP loci were located at 211 BP and 205 BP, respectively. The allele frequencies were 55% and 44 BP, respectively. The alleles of 150 BP upstream and 1 700 BP downstream were mainly located at 195 and 209 BP, while the other two loci were not concentrated. H1-H4 haploid strains were found in Laasan area of Sino-Myanmar border, and two H _ 2 haplotypes were found in three strains in Thailand. The average he of the 6 microsatellite loci in the other samples were 0.49 鹵0.230.42 鹵0.33 and 0.72 鹵0.120.67 鹵0.11.R539T, respectively, and the he at the microsatellite site adjacent to the K13 gene was significantly decreased. It was 0.17 and 0 at 150 BP upstream and 0.3 and 0 at 1 700 BP downstream, respectively. Conclusion the K13 gene of R539T mutant strain of Plasmodium falciparum is under artemisinin selection pressure in Laasan region and three regions of Thailand, and the alleles are different at 6360 BP and 31,900 BP loci upstream.
【作者單位】： 第二軍醫(yī)大學(xué)熱帶傳染病學(xué)教研室;
【基金】：國(guó)家自然科學(xué)基金(No.81220108019)~~
【分類(lèi)號(hào)】：R382.31

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