本文選題：人芽囊原蟲 + 感染情況　； 參考：《廣西醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的了解在校大學(xué)生群體人芽囊原蟲(Blastocystis hominis)的感染情況；進一步探明廣西B.h的基因型構(gòu)成,為B.h分類學(xué)研究提供科學(xué)依據(jù)。 方法采用隨機抽樣法選擇廣西醫(yī)科大學(xué)在校大學(xué)生作為流行病學(xué)調(diào)查對象,采用酸醚離心沉淀法進行糞便檢查,光鏡下形態(tài)鑒定,定性診斷。從B.h感染者糞便中分離蟲體,經(jīng)體外培養(yǎng)后抽提基因組DNA。應(yīng)用國際通用的STS引物進行PCR擴增基因分型；自行設(shè)計elf-1α保守區(qū)片段PCR引物,對廣西已發(fā)現(xiàn)的未知基因型蟲株進行DNA序列分析和同源性比較,應(yīng)用MEGA法構(gòu)建系統(tǒng)發(fā)生樹。 結(jié)果廣西醫(yī)科大學(xué)在校大學(xué)生B.h的平均感染率為14.93%；漢族感染率13.48%,壯族18.38%,其他民族感染率為17.24%；各年級感染率最低為13.19%,最高達16.28%；男性感染率為14.74%,女性感染率為15.06%；廣西本省感染率15.06%,外地生源感染率為13.94%。 以7對STS引物對53個樣本的基因組DNA進行PCR擴增,共發(fā)現(xiàn)5種已知基因型(1、3、4、6、7型)蟲株；其中,單純1型有4株,占7.55%(4/53)；單純3型17株,占全部人源性分離株的32.08%(17/53)；單純的4型有4株,占7.55%(4/53)；單純6型有1株,古1.89%(1/53)；單純7型有5株,占9.43%(5/53)；22株無特異性區(qū)帶出現(xiàn),初步認(rèn)為是未知基因型；進一步應(yīng)用elf-1α保守區(qū)引物對23株未知基因型B.h(11株來自廣西醫(yī)科大學(xué)在校大學(xué)生感染者、12株來自桂南沿海地區(qū)分離株)進行PCR擴增分析,結(jié)果有15株顯示特征性片斷,8株顯示陰性。成功進行了13株elf-1α保守區(qū)DNA測序和同源性比較分析,發(fā)現(xiàn)這13未知基因型B.h與已報道的S株系同源性達70%左右。系統(tǒng)發(fā)生樹構(gòu)建結(jié)果發(fā)現(xiàn),13株未知基因型中至少存在3種以上不同類群。 結(jié)論(1)廣西醫(yī)科大學(xué)在校大學(xué)生B.h感染率較高,并且存在校內(nèi)感染的可能性；壯族學(xué)生B.h感染率高于漢族。(2)發(fā)現(xiàn)基因1型、3型、4型、6型、7型等5種已知基因型蟲株,以3型為主；其中7型系我國首次在廣西發(fā)現(xiàn)。(3)廣西存在未知基因型B.h,初步發(fā)現(xiàn)與S株系近緣,而且存在不同類群。(4)elf-1α可作為B.h分類的另一項分類依據(jù),我們獨立設(shè)計的elf-1α保守區(qū)PCR引物能揭示未知基因型B.h之間的遺傳差異,在B.h的分類學(xué)中有應(yīng)用價值。
[Abstract]:Objective to investigate the infection of Blastocystis hominis in college students and to find out the genotypes of B.H in Guangxi, and to provide scientific basis for the study of B. h taxonomy. Methods the random sampling method was used to select the college students in Guangxi Medical University as the subject of epidemiological investigation, and the acid ether centrifugation precipitation method was used to examine the feces, the morphology was identified under light microscope, and the qualitative diagnosis was made. The genomic DNA was isolated from the feces of B. h infected individuals and cultured in vitro. The elf-1 偽 conserved fragment PCR primer was designed to analyze the DNA sequence and homology of the unknown genotypes found in Guangxi, and the phylogenetic tree was constructed by MEGA method. Results the average infection rate of B.h in college students of Guangxi Medical University was 14.933.The infection rate of Han nationality was 13.4848, that of Zhuang nationality 18.38m, that of other nationalities was 17.24; the lowest infection rate of every grade was 13.19 and the highest was 16.28m; the infection rate of male was 14.74and the infection rate of female was 15.06b; the infection rate of Guangxi was 15.06b. The infection rate in this province was 15.06 and the infection rate was 13.94. The genomic DNA of 53 samples were amplified by PCR using 7 pairs of STS primers. A total of 5 known genotypes were found, among which, 4 strains were isolated from type 1, which accounted for 7.554 / 53; 17 strains were isolated from type 3. Of all the human isolated strains, 32.08 were found in 17 / 53; 4 in type 4 alone (7.55 / 53); 1 in type 6, with 1 / 53 in ancient 1.89g / 53; 5 in type 7, accounting for 9.4355 / 53 / 22 with no specific zone, and initially considered as unknown genotypes. Furthermore, 23 unknown genotype B.h(11 strains were amplified by PCR amplification using elf-1 偽 conserved region primers from 12 strains isolated from the coastal areas of Guangxi Medical University (Guangxi Medical University). Results of the 15 strains, 8 were negative. The DNA sequencing and homology analysis of 13 elf-1 偽 conserved regions were successfully carried out. It was found that the 13 unknown genotype B.h had about 70% homology with the reported S strains. The results of phylogenetic tree construction showed that there were at least 3 different groups in 13 unknown genotypes. Conclusion (1) the infection rate of B. h in Guangxi Medical University students is higher than that in Han nationality students, and the infection rate of B.h in Zhuang nationality students is higher than that in Han nationality. The unknown genotype B.hwas found in Guangxi for the first time in China, and it was found that there is a close relationship with the S strain, and that there are different taxa, I. e., the different taxa, which can be used as another taxonomic basis for B.h classification. Our independently designed elf-1 偽 conserved region PCR primer can reveal the genetic difference between unknown genotypes B.H, and has application value in the taxonomy of B. h.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R181.3

【參考文獻】

相關(guān)期刊論文 前10條

1 李朝品,王健;淮南地區(qū)人芽囊原蟲感染的流行病學(xué)調(diào)查[J];第四軍醫(yī)大學(xué)學(xué)報;2002年16期

2 田春林,劉登宇,盧作超,劉曉泉,何登賢,趙邦權(quán);人芽囊原蟲糞便檢查與形態(tài)觀察[J];中國熱帶醫(yī)學(xué);2005年02期

3 劉常華,謝紅,鄭德福;四川省人芽囊原蟲感染情況調(diào)查[J];現(xiàn)代預(yù)防醫(yī)學(xué);1998年01期

4 金群馨,唐國都,俞開敏;人芽囊原蟲病患者腸粘膜損傷及其粘膜細胞因子的測定[J];中國寄生蟲病防治雜志;2005年05期

5 蘇子林;蘇水蓮;吳中發(fā);廖華;廖勇;謝瓊s，

本文編號：1856697