云南省昆明市HIV-1流行株<i>pol</i>基因序列特征分析

邱钰婷; 李健健; 刘家法; 王佳丽; 张米; 董兴齐

doi:10.16462/j.cnki.zhjbkz.2022.03.012

云南省昆明市HIV-1流行株pol基因序列特征分析

doi: 10.16462/j.cnki.zhjbkz.2022.03.012

邱钰婷¹,
李健健²,
刘家法²,
王佳丽²,
张米²,
董兴齐^{1, 2, ,}

1.
650000 昆明，昆明医科大学公共卫生学院
2.
650301 昆明，云南省传染病医院艾滋病关爱中心

邱钰婷和李健健同为第一作者

基金项目:

国家自然科学基金 81960605

云南省高层次卫生技术人才培养经费资助项目 H-2018050

云南省艾滋病病毒学及临床诊疗技术创新研究中心 202102AA310005

详细信息

通讯作者:
董兴齐，E-mail: ynaccdxq@126.com

中图分类号: R512.91;R181.3
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出版历程
- 收稿日期: 2021-06-07
- 修回日期: 2021-10-07
- 网络出版日期: 2022-03-17
- 刊出日期: 2022-03-10

Sequence analysis of pol gene of HIV-1 strains in Kunming, Yunnan Province

1.
Kunming Medical University, Kunming 650000, China
2.
Yunnan Provincal Infectious Diseases Hospital/AIDS Care Center, Kunming 650301, China

QIU Yu-ting and LI Jian-jian contributed equally to this article

Funds:

National Natural Science Foundation of China 81960605

Yunnan health training project of high level talents H-2018050

Yunnan AIDS virology and clinical diagnosis and treatment technology innovation research center 202102AA310005

More Information

Corresponding author: DONG Xing-qi, E-mail: ynaccdxq@126.com

摘要

摘要: 目的了解云南省昆明市人类免疫缺陷病毒1型(human immunodeficiency virus type 1，HIV-1)的亚型分布特征及其pol结构基因的变异特征。方法随机抽取2015—2019年昆明市抗病毒治疗半年以上的样本390例，收集其流行病学信息，血浆提纯HIV-1 RNA，采用巢式聚合酶链式反应扩增HIV-1 pol基因，应用Genotyping Tool、BLAST在线分型工具及MEGA 6.06软件进行基因分型，运用MEGA Distance程序计算基因距离，Entropy程序分析氨基酸多态性，进而对HIV-1流行株的pol基因序列特征进行分析。结果成功扩增获得pol基因序列318例，云南省昆明市的HIV-1亚型主要为B亚型占比3.1%，C亚型占比4.4%，CRF01_AE占比26.1%，CRF07_BC占比20.4%，CRF08_BC占比42.8%，其他亚型占比3.1%。各个亚型在性别、年龄及感染途径的构成上差异均有统计学意义(均有P < 0.001)。CRF01_AE、CRF07_BC、CRF08_BC的基因距离分别为(0.018±0.008)、(0.015±0.006)和(0.006±0.004)bp，差异均有统计学意义(均有P < 0.01)。CRF01_AE亚型中静脉药瘾感染者与性接触感染者中共有45个位点的氨基酸差异存在统计学意义(均有P < 0.05)，异性性传播与同性性传播中共156个位点有差异。结论昆明市HIV-1亚型分布复杂多样，以CRF08_BC为主，传播途径以异性性传播为主，CRF01_AE亚型为昆明地区变异程度较高的流行株，不同感染途径毒株的氨基酸突变位点存在差异，应密切监视其变化。
- 人类免疫缺陷病毒1型 /
- 亚型分布 /
- 基因距离 /
- 氨基酸变异
Abstract: Objective To investigate the distribution of human immunodeficiency virus type 1(HIV-1) subtypes and the variation of pol gene in Kunming, Yunnan Province. Methods Randomly selected 390 samples with HAART for more than half a year in Kunming from 2015 to 2019, and collected the epidemiological information. Purified the HIV-1 RNA from plasma and amplified the HIV-1 pol gene by nested polymerase chain reaction. Online analysis tools like Genotyping, BLAST and MEGA 6.06 software were used to determine strain subtypes. Distance program of MEGA was used to calculate the gene distance, and Entropy program was used to analyze the amino acid polymorphism, so as to analyze the pol gene sequence characteristics of HIV-1 strains. Results A total of 318 pol gene sequences were successfully amplified. The major subtypes of HIV-1 strains in Kunming were B subtype(3.1%), C subtype(4.4%), CRF01_AE(26.1%), CRF07_BC(20.4%), CRF08_ BC(42.8%) and other subtypes (3.1%). There were statistically significant differences in the distribution of HIV-1 genotypes in sex, age and infection routes (all P < 0.001).The genetic distances of pol gene for CRF01_AE, CRF07_BC and CRF08_BC were (0.018±0.008), (0.015±0.006) and (0.006±0.004) bp (all P < 0.01). In CRF01_ AE, there were 45 different sites in amino acid sequences between intravenous drug addicts and sexual contacts and 156 different sites between hetrosexual contacts and homosexual contacts, and the differences were sitatistically significant (P < 0.05). Conclusion There are complex and diverse HIV-1 subtypes in Kunming, and CRF08_BC is the predominant epidemic subtype. The main route of infection is heterosexual transmission. CRF01_ AE subtype is an epidemic strain with high degree of variation in Kunming. There are differences in amino acid mutation sites of strains from different infection routes, which should be closely monitored.
- Human immunodeficiency virus type 1 /
- Subtype distribution /
- Gene distance /
- Amino acid variation

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图 1 昆明市HIV-1 pol基因序列系统进化树分析

Figure 1. Phylogenetic tree analysis of HIV-1 pol gene sequence in Kunming

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图 2 CRF01_AE亚型两组不同感染途径序列氨基酸多态性分析

Figure 2. Amino acid poly morphism analysis of different infection routes in two groups of CRF01_AE subtype

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图 3 CRF01_AE亚型中异性性传播与同性性传播患者序列氨基酸多态性分析

Figure 3. Sequence amino acid polymorphism analysis of heterosexual and homosexual transmission patients in CRF01_AE subtype

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表 1 2015—2019年昆明市318例HIV/AIDS患者人口学特征及基因亚型分布情况[n(%)]

Table 1. Demographic characteristics and gene subtype distribution of 318 HIV/AIDS patients in Kunming City from 2015 to 2019 [n(%)]

变量	例数(n=318)	B	C	CRF01_AE	CRF07_BC	CRF08_BC	其他	χ²值	P值
性别								29.381	< 0.01
男	224(70.4)	9(4.0)	8(3.6)	75(33.5)	42(18.8)	82(36.6)	8(3.6)
女	94(29.6)	1(1.0)	6(6.4)	8(8.5)	23(24.4)	54(57.4)	2(2.1)
年龄(岁)								42.344	< 0.01
< 20	20(6.3)	-		2(10.0)	1(5.0)	5(25.0)	12(60.0)	-
20~ < 40	153(48.1)	6(3.9)	8(5.2)	57(37.3)	32(20.9)	42(27.5)	8(5.2)
40~ < 60	135(42.5)	4(3.0)	3(2.2)	25(18.5)	26(19.3)	75(55.6)	2(1.5)
≥60	10(3.1)	-		1(10.0)	-		2(20.0)	7(70.0)	-
感染途径								113.846	< 0.01
静脉吸毒	86(27.0)	-		3(3.5)	7(8.1)	19(22.1)	57(66.3)	-
异性性传播	120(37.3)	4(3.3)	7(5.8)	25(20.8)	25(20.8)	56(46.7)	3(2.5)
同性性传播	62(19.5)	4(6.5)	-		38(61.3)	12(19.4)	2(3.2)	6(9.7)
母婴传播	19(6.0)	-		2(10.5)	1(5.3)	5(26.3)	11(57.9)	-
其他	31(9.7)	2(6.5)	2(6.5)	12(38.7)	4(12.9)	10(32.3)	1(3.2)
合计	318(100.0)	10(3.1)	14(4.4)	83(26.1)	65(20.4)	136(42.8)	10(3.1)
注：其他亚型(例数)依次为A(2)、CRF55_01B(6)、CRF78_cpx(1)以及CRF86_BC(1)。

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表 2 2015—2019年昆明市318例HIV/AIDS患者HIV-1 pol基因的三种主要亚型基因距离

Table 2. Gene distances of three main subtypes of HIV-1 pol gene in 318 HIV/AIDS patients in Kunming from 2015 to 2019

HIV-1亚型	例数(n=276)	基因距离(x±s, bp)	F值	P值
CRF01_AE	75	0.018±0.008	10 739.856	< 0.001
CRF07_BC	58	0.015±0.006
CRF08_BC	143	0.006±0.004

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