2010-2011年广州市白云区某社区流动人口病毒性腹泻的监测

发布时间：2018-02-08 11:18

  本文关键词： 腹泻 轮状病毒 诺如病毒 腺病毒 星状病毒 流动人口 监测　出处：《南方医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：研究背景： 感染性腹泻是人类最常见的疾病之一,是由细菌、病毒、原虫等多种病原体所引起的、以腹泻症状为主并广泛存在并流行于世界各地的肠道传染病,也是当今全球性重要的公共卫生问题,位居三大传染病死亡原因之一,尤其在儿童中更是一种常见病和多发病,是导致儿童死亡的两大病因之一。特别在发展中国家,腹泻是导致儿童死亡的主要原因。《2003年世界卫生报告》中披露,发展中国家感染性腹泻导致儿童死亡达156.6万人,占死亡人数的15.2%,居死因第三位。在我国感染性腹泻的发病率居所有传染病之首。目前,病毒已成为感染性腹泻的重要病原体。1972年发现诺瓦克病毒后,已相继发现和确立引起腹泻的病毒有呼肠孤病毒科(轮状病毒)、腺病毒(肠腺病毒)、杯状病毒科(诺如病毒和扎幌样病毒)、星状病毒科(星状病毒)和冠状病毒科的成员。 轮状病毒(Rotavirs, RV)是婴幼儿腹泻的最主要病因之一,根据抗原性的不同将轮状病毒分为7个组(A-G)。轮状病毒感染常无症状,有症状感染的潜伏期为1-3天,病程常5-7天,主要临床表现是发热、频繁的呕吐、水样腹泻、轻度失水和酸中毒。据报道,RV肠炎好发于6-24个月的婴幼儿。在气候随四季变化的地区,RV感染的高峰季节随时间和区域不同而有所变动,多发生在寒冷干燥的秋冬季节,在终年炎热的地区全年均可发生,亚热带地区婴幼儿RV腹泻多发生于秋冬季,流行高峰在10-12月,而春季3-5月份又可有一小高峰,突然的环境改变亦可能影响流行,世界各地均有散发和流行的报道。 国内外大量研究表明,诺如病毒(Norovirus, NVs)是仅次于RV导致人类急性非细菌性胃肠炎(acute gastroenteritis, AGE)的重要病原体,NVs主要通过粪-口传播,是导致食源或水源性急性腹泻的主要病原,世界范围内引起低龄儿童急性病毒性胃肠炎的主要病原,同时也是造成包括学校、家庭、旅游区、医院、食堂、军队的爆发性胃肠炎流行的重要病原,对社会生产力产生巨大破坏力,其地位仅次于RV。该病毒的特点是发病率高、致病剂量低和抵抗力强。 腺病毒(Adenovirus, AdV)在儿童腹泻标本中的检出率虽不如RV高,但近两年随着对AdV的了解越来越深入,检测方法不断增加,商品化的检测试剂盒越来越快速简便,各国对AdV引起的儿童腹泻越来越重视。 世界大部分地区均有星状病毒(Astrivirus, AstV)感染的报道,已证实它是引起婴幼儿、老年人及免疫低下者腹泻的重要病原之一。资料显示AstV成为婴幼儿病毒性胃肠炎的第2或第3位病原体。AstV感染常见于秋冬季,多为散发,亦可在医院、幼儿园、老人院、学校和军队引起爆发流行。 然而,随着我国城市化进程和经济建设的发展,大量外来人口融入大城市,使得城乡结合部、城中村等经济条件相对落后、人员构成复杂、流动性大的地区腹泻病的情况不容乐观。由于城市流动人口存在流动性强、知识水平低、经济条件差等问题,整体上影响了城市流动人口的卫生保健服务状况。 研究目的： 1.了解广州市某社区2010-2011年流动人口病毒性腹泻RV、NVs、AdV和AstV感染状况、临床表现及流行病学特征； 2.了解该地区病毒性腹泻中NVs、AdV和AstV基因型别,探讨流动人口中三种病毒的流行优势毒株； 3.丰富南方病毒感染性腹泻的流行病学资料,为流动人口病毒性腹泻的防治提供一定的参考依据。 研究方法： 1.标本来源：收集2010年5月～2011年12月期间在广州市白云区某社区卫生服务中心就诊的383例流动人口腹泻患者粪便标本、人口学资料、临床及流行病学资料。 2.标本处理方法：每份粪便标本2g,分别各加2mlPBS,震荡混匀,分装后-80℃保存备用。 3.标本病毒检测方法：利用RT-PCR和PCR方法,用引物JV12Y/JV13I、 AD1/AD2和Mon269/Mon270分别对所有标本进行NVs、AdV和AstV感染的检测,对PCR阳性产物进行纯化回收、测序,应用BLAST将测序结果在GenBank上寻找各阳性株的同源序列,使用MEGA5.0生物软件进行序列的相似性比对,分析NVs、AdV和AstV的基因型别,判断NVs、AdV和AstV的流行优势毒株。 4.统计分析方法：运用统计软件SPSS13.0对收集的患者资料进行统计分析,采用描述性分析,计数资料单因素分析采用卡方检验,多因素分析运用logistic逐步回归,检验水准α为0.05。 结果： 1.人口学资料：2010年5月～2011年12月共收集广州市某社区腹泻患者粪便标本和问卷共383分。检测情况：5岁以下儿童占90.9%(348/383),来自广东省(除广州市外)其他地区、湖南和江西等周边地区。流动儿童其监护人文化水平较低,以中学水平为主,占89.7%(323/383),多数监护人为无业。 2.病毒性腹泻流行病学特征：病毒性腹泻在监测期内均可发生,于秋冬季节病例较多,感染高峰出现在10、11月,但8月出现小高峰；4种病毒的阳性病例为214例,检出率为55.9%(214/383),混合感染率为10.4%(40/383)；主要感染5岁以下儿童,占94.4%(202/214)；其中,男性患者139例,女性75例,男女患者比例为1.85：1。 3. RV、 NVs、AdV和AstV的检测结果和流行病学特征：RV、NVs、AdV和AstV的检出率分别为43.1%(165/383)、10.7%(41/383)、7.0%(27/383)和6.0%(23/383),四种病毒阳性检出者年龄均主要集中在5岁以下,年龄组间和性别间各种病毒的检出情况差异无统计学义,RV感染高峰发生在秋冬季,NVs和AdV的高峰均显示在温度较高的夏季,而AstV在2010年5月-2010年12月期间,8月检出率最高,2011年全部AstV感染均发生在秋冬季(10月-12月)。 4.无病毒检出组(A组)和病毒检出组(B组)相关分析：单因素分析显示,A和B两组在患者职业(Fisher检验,P=0.032)和文化程度(Fisher检验,P=0.034)方面差异有统计学意义,表明感染性腹泻主要感染学龄前儿童；两组的粪便量()(χ2=5.496,P=0.019)、粪便性状(Fisher检验,P=0.007)、腹泻方式(χ2=9.296,P=0.026)、食欲(χ2=11.968,P=0.001)及乏力(χ2=6.647,P=0.010)情况等临床表现方面差异有统计学意义；而其他人口学资料、临床表现和流行病学接触史等方面差异无统计学意义。 5. NVs、AdV和AstV的序列分析结果：383份粪便标本中,共检出NVs41例,系统进化树和相似性分析显示：以GII-4型为主(38例),其中35株为GII.4-2006b亚型、2株为GII.4-2006a亚型、1株为经典GII-4型,另外分别为GII-6、GⅡ-14和GI-2型；检出27株AdV,系统进化和相似性分析显示：16株为AdV41型,其中2株为AdV41型GTC1型,14株为AdV41型GTC2型,AdVl、2和12各检出两株,AdV3和AdV7各检出1株,AdV31检出3株；而检出23株AstV均为AstV-1. 结论： 1.监测情况显示：腹泻患者或患者家属(14岁以下患儿家属)文化层次较低,以中学文化水平为主,符合流动人口整体文化程度较低的趋势。 2.病毒性腹泻的检出率为55.9%(214/383),该病全年均有散发,发病高峰在秋冬季,但是夏季有小高峰出现；两种及两种以上病毒混合感染较常见,检出率为10.4%,患者主要是2岁以下婴幼儿。混合感染是否会加重病情,是应该受到关注的问题。 3.病毒为流动儿童腹泻的重要病因,其中轮状病毒是最主要的病原体,其次依次为NVs、 AdV和AstV。 RV以秋冬季高发；NVs夏季为感染高峰,NVs GⅡ.4基因型为广州流动人口NVs感染的流行优势毒株,与我国国内情况相同。其中变异株GII.4-2006b38株、GII.4-2006a2株,而GII.4-2006b曾引起2007年后世界范围内多个国家腹泻疫情大量增加,我国较多地区均已检测到2006b,但未见2006a相关报道,本次研究检出两株NVs2006a引起的腹泻,显示2006a亚型在我国境内也有发病,应增加相关研究。此次研究中夏季较多AdV感染,表明AdV41型仍为我国腺病毒感染腹泻的主要流行毒株,同时存在其他多个AdV型别的散在感染,但并未见AdV40型感染。2010年AstV感染主要发生在夏季,而2011年则全部发生在秋冬季节,AstV阳性均为AstV-1型,显示AstV-1为广州流动人口AstV感染的流行优势型别。 4.无病毒检出组(A组)和病毒检出组(B组)在患者职业和文化程度方面差异有统计学意义,说明主要为学龄前儿童,显示腹泻病原体主要感染年龄较小儿童；两组的粪便量、粪便性状、腹泻方式、食欲及乏力情况等临床表现方面差异有统计学意义,表现为病毒检出组患者粪便量较无检出组偏多,主要为水样便或蛋花样便,并对患者食欲影响较大,伴有乏力,因此对病毒感染患者尽早进行对症支持治疗,防止因失水而加重病情。
[Abstract]:Research background:
Infectious diarrhea is one of the most common diseases of mankind, is caused by bacteria, viruses, parasites and other pathogens caused by diarrhea, intestinal infectious disease symptoms and are widespread and popular in the world, is also the important global public health problem, is one of the causes of three infectious diseases, especially in children it is a common and frequently occurring disease, is the two leading cause of death in children, especially in developing countries, resulting in disclosure.<2003 diarrhea is the leading cause of death among children in the World Health Report "in developing countries, infectious diarrhea in children killed up to 1 million 566 thousand people, accounting for 15.2% of the number of deaths, the leading cause of death in third. The incidence of infectious diarrhea in China's first rate in all infectious diseases. At present, the virus has become the important pathogens of infectious diarrhea by.1972 years after the virus has been found in Novak, and found the The establishment of diarrhea caused by the virus Reoviridae (rotavirus), adenovirus (enteric adenovirus), Caliciviridae (norovirus and Sapporo), astrovirus (astrovirus) and members of the coronaviridae family.
Rotavirus (Rotavirs, RV) is one of the main causes of infantile diarrhea, according to the antigenicity of different rotavirus were divided into 7 groups (A-G). Rotavirus infection is often asymptomatic, symptomatic infection incubation period is 1-3 days, the duration of the disease was 5-7 days, the main clinical manifestations were fever, frequent vomiting, watery diarrhea, mild dehydration and acidosis. According to reports, RV enteritis occurs in 6-24 months infants. In the regional climate changes with the seasons, the peak season of RV infection with time and area vary, occurred in the cold and dry autumn winter festival, occurred in the area year-round hot throughout the year may, subtropical infant RV diarrhea occur in autumn and winter, the epidemic peak in 10-12 months, and 3-5 months in spring may have a small peak, sudden environmental change may also affect the popular all over the world, sporadic and epidemic reports.
A large number of domestic and foreign research shows that norovirus (Norovirus, NVs) is next to the RV cause of acute nonbacterial gastroenteritis in humans (acute gastroenteritis, AGE) an important pathogen, NVs mainly through the fecal oral transmission is the main food source disease or waterborne acute diarrhea, the main pathogen of children acute viral gastroenteritis worldwide, but also causes include schools, families, tourist areas, hospitals, canteens, an important pathogen of military explosive epidemic gastroenteritis, produce great damage to social productivity, the characteristics of its position after the RV. of the virus is a high incidence rate, low dose of pathogenic and strong resistance.
Although the detection rate of Adenovirus (AdV) in children's diarrhoea specimens is not as high as that of RV, in recent two years, with the increasing understanding of AdV, the detection methods are increasing. Commercialized detection kits are more and more fast and simple. All countries pay more and more attention to the diarrhea caused by AdV.
Have astrovirus in most parts of the world (Astrivirus, AstV) infection reports, has confirmed that it is one of the most important pathogens causing infant, the elderly and immunocompromised diarrhea. Data show that AstV of infant viral gastroenteritis second or third pathogen.AstV infection is common in autumn and winter, more distributed, but also at the hospital. Kindergarten, homes, schools and army outbreaks.
However, with the development of China's city development and economic construction, a large number of foreign population into the city, making the urban and rural areas, villages and other economic conditions are relatively backward, in a complex flow area of diarrhea of the big city. The situation is not optimistic because of the existence of floating population mobility, the low level of knowledge, problem poor economic conditions, the overall effect of the health care service of floating population in the city.
The purpose of the study is:
1. to understand the infection status of RV, NVs, AdV and AstV, clinical manifestations and epidemiological characteristics of viral diarrhea in a community of 2010-2011 years in a community of Guangzhou.
2. to understand the genotypes of NVs, AdV and AstV in viral diarrhea in this area, and to explore the dominant strains of three viruses in the floating population.
3. to enrich the epidemiological data of infectious diarrhea of the southern virus, and to provide some reference for the prevention and treatment of viral diarrhea of the floating population.
Research methods:
1. source of specimens: collected 383 stools, demographic data, clinical and epidemiological data of floating population diarrhoea patients in a community health service center in Baiyun District of Guangzhou from May 2010 to December 2011.
2. sample treatment method: each faecal specimen 2G, each adding 2 mlPBS respectively, concussion and mixing, after loading, -80 centigrade preservation.
Methods 3. specimens of virus detection: using RT-PCR and PCR methods, using primers JV12Y/JV13I, AD1/AD2 and Mon269/Mon270 respectively by NVs for all the samples, the detection of AdV and AstV infection, the positive PCR products were purified and sequenced by BLAST sequencing results in GenBank to find the homologous sequences of positive strains, similarity comparison the sequence using the MEGA5.0 biological analysis software NVs, AdV and AstV genotypes, NVs AdV and AstV of the judgment, the prevailing strain.
4. statistical analysis: statistical software SPSS13.0 was used to analyze the collected data. Descriptive analysis, counting data, single factor analysis, chi square test, multivariate analysis and logistic stepwise regression were used to test the level of alpha 0.05..
Result锛，

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