冬春季儿童、婴幼儿哮喘急性发作的呼吸道病毒病原学特征及临床研究

目的 了解北京地区冬春季儿童、婴幼儿哮喘急性发作时的呼吸道常见病毒——呼吸道合胞病毒（RSV）、流感病毒（IFA1、IFA3、IFB）、副流感病毒（PIF1、PIF3）、腺病毒（ADV）的感染情况及与临床症状和嗜酸细胞的关系。方法 对2000年11月至2001年3月及2001年11月至2002年3月，来首都儿科研究所哮喘中心就诊的哮喘急性发作的患儿176名，采用病毒分离及间接免疫荧光法，对鼻咽分泌物（nasopharyngeal secretions，NPS）中七种病毒抗原进行监测，并同时记录临床症状和用药情况及进行鼻咽分泌物和外周血中嗜酸细胞计数。结果 176例哮喘急性发作患儿NPS中，79例检测出病毒，阳性率为44．9％。其中RSV 66例，感染率为37．5％、IF7例（4．0％）、ADV6例（3．4％）及PIF4例（2．3％）。RSV占79例病毒感染患儿的83．5％；4例患儿同时测定出RSV和其他病毒混合感染。这些病毒在哮喘急性发作的患儿中检出率与年龄呈反比，小年龄组的患儿病毒感染多；检出病毒的患儿病情重，伴发热的患儿显著多于未检查出病毒的患儿。病毒感染与非感染组的NPS中嗜酸细胞数目检测无明显差别；但血中嗜酸细胞的数目，病毒检出阳性的患儿，较病毒测定阴性的患儿明显减少（t=2．676，P〈0．001）。结论 北京地区冬季近半数哮喘急性发作的患儿呼吸道病毒检测阳性，其中RSV是婴幼儿哮喘发作的主要感染病毒，引起较严重的临床症状；病毒检出阳性的患儿，血中嗜酸细胞的数目明显减少。     

Viral diarrhea in children in Beijing, China

A study was undertaken from November 1994 to August 1996 to determine the role of viruses in children (< or =5 years of age) hospitalized at Beijing Children Hospital, Beijing China, for acute diarrhea. Stool samples from diarrheal patients were investigated by ELISA, electron microscopy, and RT-PCR for the presence of rotavirus, calicivirus, astrovirus, and adenovirus. Group A rotavirus was detected in 55.9% of all diarrheal patients and comprised 82.5% of all viruses detected. Group A rotavirus samples were further characterized for their G-type specificity by RT-PCR. Four major G types (1-4) were identified. G1 to G4 accounted for 58.9%, 15.7%, 16.8%, and 6.3%, respectively, of the serotyped samples. Almost all rotavirus infections occurred in children less than 1 year of age, with a significant clustering during the winter months. Group C rotavirus was detected in one 18-month-old child. Astroviruses, caliciviruses, and adenoviruses were detected in 8.5%, 7.6%, and 2.5% of the hospitalized children, respectively. This, the first viral etiological study of childhood diarrhea in China, concludes that rotavirus G1-4 strains play an important role in severe diarrhea in Beijing children.     

珠海市冬春季节儿童病毒性腹泻的流行病学特征

目的 探讨珠海市冬春季节儿童诺如病毒(NV)、扎如病毒(SV)和星状病毒(AstV)等3种重要病毒性腹泻的流行病学特征.方法 采集2009年11月21日至2010年4月3日在珠海市妇幼保健院就诊的腹泻儿童粪便样本,选取轮状病毒和腺病毒筛查阴性的样本,采用逆转录-聚合酶链反应(RT-PCR)技术检测NV、SV和AstV的特异基因片段,并对诺如病毒检测阳性的样本进行分子分型.结果 3种病毒季节总检出率为21.49％,其中2009年12月的检出率最高,为29.05％,而2010年2月检出率最低,为12.20％,87.96％的阳性样本来自0～30月龄患儿.NV、SV及AstV的季节检出率分别为14.70％、2.75％和4.04％,3种病毒中以NV和SV各月份阳性检出率差别较大,而AstV较为一致,12 ～ 18月龄患儿NV检出率最高(34.09％),60 ～ 120月龄患儿SV检出率最高(12.5％),24 ～30月龄患儿AstV检出率最高(16.67％).诺如病毒经分子分型后均为GⅡ型.结论 NV是引起2009年冬春季节珠海市儿童病毒性腹泻的主要病原之一,SV与AstV也是重要的病原,应加强对婴幼儿NV、SV和AstV等3种重要病毒性腹泻的监测和防控工作.     

成都地区婴幼儿病毒性腹泻分子流行病学研究

目的 通过分子流行病学研究成都地区婴幼儿病毒性腹泻的病原学特点,掌握本地区病原分布特征,为疫苗研制和疫情控制提供科学依据.方法 采用酶联免疫吸附试验( ELISA)及逆转录聚合酶链反应(RT-PCR)对成都地区2006 -2008年度376例婴幼儿腹泻粪便标本进行轮状病毒(RV)、杯状病毒(HuCV)、星状病毒(AstV)及肠道腺病毒(Adv)检测.结果 RV的检出率为37.76％(142/376),其中42例进行G分型,45例进行P分型,G分型以G3型为主21例(50％),其次为G2和G1型,P分型以P[8]型为主21例,其次是P[4]型19例.RV感染主要为6～23月的婴幼儿,发病高峰在10 ～12月份(75.8％).RT-PCR法检出HuCV、AstV及Adv的检出率分别为15.85％、1.64％及2.04％.结论 RV是成都地区婴幼儿病毒性腹泻的主要病原,其流行的主要血清型为G3型、P[8]、P[4]型.除RV外,HuCV也是重要的病原.     

成都地区婴幼儿病毒性腹泻分子流行病学研究

目的通过分子流行病学研究成都地区婴幼儿病毒性腹泻的病原学特点,掌握本地区病原分布特征,为疫苗研制和疫情控制提供科学依据。方法采用酶联免疫吸附试验（ELISA）及逆转录聚合酶链反应（RT—PCR）对成都地区2006—2008年度376例婴幼儿腹泻粪便标本进行轮状病毒（RV）、杯状病毒（HuCV）、星状病毒（AstV）及肠道腺病毒（Adv）检测。结果RV的检出率为37．76％（142／376）,其中42例进行G分型,45例进行P分型,G分型以G3型为主21例（50％）,其次为G2和G1型,P分型以P[8]型为主21例,其次是P[4]型19例。RV感染主要为6～23月的婴幼儿,发病高峰在10—12月份（75．8％）。RT—PCR法检出HuCV、AstV及Adv的检出率分别为15．85％、1．64％及2．04％。结论RV是成都地区婴幼儿病毒性腹泻的主要病原,其流行的主要血清型为G3型、P[8]、P[4]型。除RV外,HuCV也是重要的病原。     

Identification of viral agents causing gastroenteritis among children in Buenos Aires, Argentina.

BACKGROUND: Gastroenteritis is one of the most common diseases affecting children and rotavirus is the major etiological agent worldwide. Although the role of rotaviruses as a causal agent of gastroenteritis has been previously established in Argentina, little is known about the frequency of other gastrointestinal viruses. OBJECTIVES: The present study was designed to determine the frequency of calicivirus in comparison with rotavirus, astrovirus and adenovirus type 40/41 in our country. STUDY DESIGN: Sixty six diarrhea patients (10%) =3 years of age, were randomly selected from a previous series (9/1997-8/1998) and virus frequency determined using RT-PCR and/or commercial EIAs. In addition, we sequenced the RNA polymerase region of calicivirus-positive samples to characterize Argentine strains. RESULTS: We detected at least one viral agent in 48.5% (32/66) of the patients. Rotavirus, calicivirus, astrovirus, and enteric adenovirus were found in 17, 16, 5, and 2 children, respectively. Sequence analysis of 4 calicivirus positive samples allowed us to identify a strain clustered to Lorsdale virus, one strain clustered with Manchester virus and two strains clustered with London/92 virus. CONCLUSIONS: Our data suggest that calicivirus and rotavirus are major agents of gastroenteritis in children younger than 3 years of age in Buenos Aires, Argentina. Unfortunately, viral diagnosis is seldom available in Argentine hospitals and coproculture results are mostly negative. Our data supports the need to develop rapid and sensitive tests for calicivirus diagnosis. The introduction of affordable viral diagnosis in our pediatric hospitals will improve patient care by reducing the unnecessary use of antibiotics.     

Etiology of childhood diarrhea in Beijing, China.

To determine the role of recently recognized enteropathogens in childhood diarrhea in China, 221 children with diarrhea and 108 controls seen at the Beijing Children's Hospital were studied during April and May 1989. Stools were examined for ova, parasites, and rotavirus, cultured for bacterial pathogens, and probed for enterotoxigenic Escherichia coli (ETEC), enteroinvasive E. coli (EIEC), enterohemorrhagic E. coli (EHEC), and enteropathogenic adherence factor-positive (EAF+) E. coli. Pathogens were identified in 56.5% of children with diarrhea and 43.5% of controls (P = 0.04). Detection of enteropathogens was significantly greater in patients examined within 1 week of symptom onset (65%) than in patients examined later (39%; P = 0.01). ETEC was the most frequently detected pathogen in children with diarrhea, accounting for 20% of the cases. Other agents identified in patients included the following: salmonellae, 12%; rotavirus, 7%; EIEC, 7%; EHEC, 7%; members of the Aeromonas hydrophila group, 6%; EAF+ E. coli, 5%; Ascaris lumbricoides, 3%; shigellae, 3%; campylobacters, 2%; and Vibrio spp., 0.5%. The isolation rates of salmonellae (P = 0.02), EAF+ E. coli (P = 0.04), and mixed pathogens (P = 0.05) were significantly greater for diarrhea patients than for controls. Resistance to multiple antimicrobial agents occurred in 39% of the Salmonella isolates, 22% of the Aeromonas isolates, and 17% of the Shigella isolates. Multiresistant salmonellae (P = 0.05) and shigellae were recovered from diarrheal stools only. Ciprofloxacin, cefotaxime, and imipenem were the only agents tested to which all bacterial isolates were susceptible in vitro. These results suggest that both traditional and newly recognized agents are important causes of childhood diarrhea in Beijing and that therapy may be complicated by indigenous antimicrobial resistance.     

Parasitic, bacterial, and viral enteric pathogens associated with diarrhea in the Central African Republic.

A total of 1,197 diarrheic children less than 15 years old were investigated for parasitic, bacterial, and viral enteropathogens from March 1981 through February 1982 in the Central African Republic. One or more pathogens were identified from 49.4% of the patients. Rotavirus was the most frequently identified pathogen among children less than 18 months old. Enteropathogenic Escherichia coli was the second most frequently isolated pathogen (12.1%) in children less than 2 years of age. Campylobacter jejuni was also isolated frequently from diarrheic children less than 5 years of age (10.9%). Entamoeba histolytica was identified in very young children and was found to be the most frequent enteropathogen associated with diarrhea in children over the age of 2 years. Enterotoxigenic Escherichia coli was rarely isolated (ca. 2%). There was a peak in the incidence of rotavirus during the dry season and in the incidence of Campylobacter jejuni during the rainy season.     

Rotavirus serotype G5 associated with diarrhea in Brazilian children.

Rotavirus serotype G5 in fecal specimens of 38 Brazilian children with diarrhea was identified by PCR and enzyme immunoassays. The strains exhibited long RNA electropherotypes and either subgroup II or nonsubgroup I-nonsubgroup II specificities. Serotype G5 has been found in piglets and horses but not yet in humans.     

The use of multiplex PCR for the diagnosis of viral severe acute respiratory infection in children: a high rate of co-detection during the winter season

Respiratory tract infection is a major cause of hospitalization in children. Although most such infections are viral in origin, it is difficult to differentiate bacterial and viral infections, as the clinical symptoms are similar. Multiplex polymerase chain reaction (PCR) methods allow testing for multiple pathogens simultaneously and are, therefore, gaining interest. This prospective case-control study was conducted from October 2013 to February 2014. Nasopharyngeal (NP) and oropharyngeal (throat) swabs were obtained from children admitted with severe acute respiratory infection (SARI) at a tertiary hospital. A control group of 40 asymptomatic children was included. Testing for 16 viruses was done by real-time multiplex PCR. Multiplex PCR detected a viral pathogen in 159/177 (89.9 %) patients admitted with SARI. There was a high rate of co-infection (46.9 %). Dual detections were observed in 64 (36.2 %), triple detections in 17 (9.6 %), and quadruple detections in 2 (1.1 %) of 177 samples. Seventy-eight patients required intensive care unit (ICU) admission, of whom 28 (35.8 %) had co-infection with multiple viruses. AdV, HBoV, HRV, HEV, and HCoV-OC43 were also detected among asymptomatic children. This study confirms the high rate of detection of viral nucleic acids by multiplex PCR among hospitalized children admitted with SARI, as well as the high rate of co-detection of multiple viruses. AdV, HBoV, HRV, HEV, and HCoV-OC43 were also detected in asymptomatic children, resulting in challenges in clinical interpretation. Studies are required to provide quantitative conclusions that will facilitate clinical interpretation and application of the results in the clinical setting.     

北京市东城区5岁以下婴幼儿病毒性腹泻病原学研究

目的 了解北京市东城区5岁以下婴幼儿病毒性腹泻的主要病原体.方法 2011年10月-2015年9月在东城区第二妇幼保健院收集1 754例5岁以下腹泻患儿便标本,用RT-PCR和PCR法检测轮状病毒、诺如病毒、札如病毒、星状病毒和肠道腺病毒.结果 标本腹泻病毒总检出率为36.0％(632/1 754);轮状病毒检出率12.4％;诺如病毒检出率10.1％;札如病毒检出率5.4％;肠道腺病毒检出率9.4％;星状病毒检出率3.5％.腹泻病毒检出高峰为每年10月-次年2月.诺如病毒和肠道腺病毒以24～60月龄患儿检出率较高,其余几种病毒以6～24月龄检出率较高;男女性别检出率没有统计学差异(x2=0.276,P＞0.05).结论 婴幼儿腹泻病人中5种病毒均有检出,其中轮状病毒和诺如病毒是主要的病原体,肠道腺病毒检出率明显高于国内其他地区.     

Etiology and epidemiology of viral diarrhea in children under the age of five hospitalized in Tianjin, China

Viral diarrhea is a great threat to children's health in developing countries. We conducted a prospective surveillance study of acute diarrhea of young children at Tianjin Children's Hospital from April 2008 to April 2009. Viral infections were detected in 356 of the total 766 collected stool specimens (46.48%). Rotavirus infections were the most common (27.94%; predominant type G1), followed by adenovirus infections (17.62%; predominant type Ad41), norovirus infections (5.87%; predominant type GII-4/2006 b), and astrovirus infections (3.15%; only HAstV-1). Children younger than 1 year old were the most susceptible population to viral infections (87.9%). Diarrhea, vomiting, and fever were the most frequent clinical symptoms among the infected patients. The viral infections had no age, sex, or regional differences. Most infection rates were higher in the autumn, winter, and spring. This study supported that the rotavirus vaccine should be included in the Expanded Programme on Immunization in China.     

Human reovirus-like agent as the major pathogen associated with "winter" gastroenteritis in hospitalized infants and young children.

We found a human reovirus-like agent in the stools of 42 per cent of 143 infants and young children hospitalized with acute gastroenteritis between January, 1974, and June, 1975. Half the patients studied by electron microscopy and serologic technics had evidence of infection with the agent. The infection had a seasonal pattern: 59 per cent of those admitted during the cooler months (November to April) shed the agent, with a peak of 78 per cent in December, 1974, and January, 1975, combined. None of the patients admitted during the warmer months (May to October) shed the agent. None of 275 Escherichia coli isolates from 32 patients with diarrhea produced heat-labile enterotoxin, whereas 17 of the 32 had evidence of infection with the reovirus-like agent. In addition, 14 of 40 parents of 37 patients with diarrhea associated with the reovirus-like agent were also infected, but most infectious were inapparent. This agent appears to be the major cause of diarrheal illness in the young during the cooler months.     

Etiology of diarrhea in young children in Denmark: a case-control study

Infectious gastroenteritis is one of the most common diseases in young children. To clarify the infectious etiology of diarrhea in Danish children less than 5 years of age, we conducted a 2-year prospective case-control study. Stools from 424 children with diarrhea and 870 asymptomatic age-matched controls were examined, and their parents were interviewed concerning symptoms. Rotavirus, adenovirus, and astrovirus were detected by enzyme-linked immunosorbent assay, and norovirus and sapovirus were detected by PCR. Salmonella, thermotolerant Campylobacter, Yersinia, Shigella, and Vibrio spp. were detected by standard methods. Shiga toxin-producing (STEC), attaching-and-effacing (A/EEC), enteropathogenic (EPEC), enterotoxigenic, enteroinvasive, and enteroaggregative Escherichia coli were detected by using colony hybridization with virulence gene probes and serotyping. Parasites were detected by microscopy. Overall, a potential pathogen was found in 54% of cases. More cases than controls were infected with rotavirus, Salmonella, norovirus, adenovirus, Campylobacter, sapovirus, STEC, classical EPEC, Yersinia, and Cryptosporidium strains, whereas A/EEC, although common, was not associated with illness. The single most important cause of diarrhea was rotavirus, which points toward the need for a childhood vaccine for this pathogen, but norovirus, adenovirus, and sapovirus were also major etiologies. Salmonella sp. was the most common bacterial pathogen, followed by Campylobacter, STEC, Yersinia, and classical EPEC strains. A/EEC not belonging to the classical EPEC serotypes was not associated with diarrhea, underscoring the importance of serotyping for the definition of EPEC.     

Identification and characterization of a novel subgenotype of bovine viral diarrhea virus isolated from dairy cattle in Northwestern China

Bovine viral diarrhea virus (BVDV) was detected by RT-PCR in 105 out of 391 samples which were collected from five dairy farms in Ningxia, China during 2010–2011. Non-cytopathogenic BVDV was isolated from 13 samples and a 230-bp fragment of the 5′-untranslated region was amplified and sequenced. While the predominant subgenotypes were BVDV-1b and BVDV-1d, a potentially novel subgenotype was identified by phylogenetic analysis, which may have implications for vaccine development.     

Characterization of a bovine viral diarrhea virus originated from cattle in Gansu Province, China

A bovine viral diarrhea disease virus (BVDV) GS-4 was isolated in Western China form dairy cattle with respiratory disease. Genomic comparison analysis with the 5′ half genome sequence encompassing the coding region of N^pro, capsid, and envelope glycoproteins showed that the GS-4 should be classified into BVDV-1b1, which is considered as one of the predominant subgenotypes found in China. This classification was confirmed by phylogenetic analysis based on E2 coding region.     

Tubular structures associated with acute nonbacterial gastroenteritis in young children.

We clearly demonstrated a fine morphology of the tumbular structures antigenically related to the reovirus-like particles associated with acute nonbacterial gastroenteritis in young children. The tubular structures with a caplike structure on each end seemed to be complete forms, 75 to 80 nm in width and approximately 1,000 nm in length.     

广州流动儿童病毒性腹泻监测

目的了解广州流动儿童病毒性腹泻的感染情况。方法收集2010年5月至2011年4月广州市白云区某社区卫生服务站241例流动儿童腹泻患者的人口资料学以及粪便标本,利用酶联免疫技术（ELISA）检测轮状病毒（RV）,采用逆转录聚合酶链反应（RT-PCR）法检测标本中诺如病毒（NVs）、星状病毒（AstV）,聚合酶链反应（PCR）法检测腺病毒（AdV）。结果 241例患者粪便标本中RV、NVs、AdV、AstV4种病毒总检出率为56.0%（135/241）,各病毒检出率分别为43.6%（105/241）、11.2%（27/241）、7.9%（19/241）、6.6%（16/241）。27株NVs阳性毒株均为GII-4;19株AdV中14株为AdV-41型,并有AdV1、2、3、31型的散在感染;AstV感染15株,均为AstV-1型。结论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,其他依次为NVs、AdV和AstV。