Acute gastroenteritis (AGE) in children can be attributed to a multitude of bacterial and viral pathogens. The objective of this study was to investigate the epidemiology of bacterial and viral AGE in children and to compare clinical characteristics between single and multiple enteric pathogen infections. A total of 456 stool samples were collected from outpatient children under 5 years old with AGE, which were subsequently analyzed for nine bacteria and three viruses using the Luminex xTAG® Gastrointestinal Pathogen Panel. The presence of at least one pathogen was detected in 260 cases (57.0%), with Salmonella being the predominant agent, followed by norovirus, Campylobacter, and rotavirus. A total of 69 cases (15.1%) exhibited positive results for two or more enteric pathogens. Although certain co-infections demonstrated significant differences in primary clinical features compared with mono-infections, no statistical variance was observed in terms of disease severity. In outpatient children from southern China, Salmonella emerged as the most prevalent causative agent of AGE, succeeded by norovirus and Campylobacter. This study underscores the burden posed by coinfections and highlights the clinical characteristics associated with AGE when accompanied by coinfections among children under 5 years old.

目的 建立一种检测登革病毒(dengue virus,DENV)特异性抗体的双抗原夹心ELISA方法.方法 利用毕赤酵母系统表达4个血清型登革病毒包膜蛋白Ⅲ区(envelope protein domain Ⅲ,ED Ⅲ),并采用改良过碘酸钠法对EDⅢ蛋白标记辣根过氧化物酶(Horseradish peroxidase,HRP),建立一种可同时检测4个血清型登革病毒ED Ⅲ蛋白特异性抗体的双抗原夹心ELISA法.并对2014年广州珠江医院门诊和住院确诊的登革热患者血清标本进行检测,并与澳洲Panbio MAC ELISA检测的敏感性进行比较.结果 本研究成功建立了一种检测4个血清型登革病毒EDⅢ蛋白特异性抗体的双抗原夹心ELISA法,该方法能同时检测到4个血清型登革病毒ED Ⅲ蛋白免疫的小鼠血清和Ⅰ型登革病毒EDⅢ蛋白免疫的兔血清,而与烟曲霉AF-MP蛋白免疫的小鼠血清和兔血清均无交叉反应.对184份健康人血清标本检测全为阴性,而对168份确诊为登革病毒感染患者血清标本检测结果表明,两种诊断方法检测结果差异无统计学意义,P＜0.001(57.1％vs 57.7％),联合NS1抗原检测方法,其敏感性提高到97.6％.结论 双抗原夹心ELISA法检测登革病毒ED Ⅲ特异性抗体具有高度的特异性,但如果能同时联合抗原检测可明显提高登革热的早期诊断率.

Abstract Background Human astrovirus (HAstV) and sapovirus (SaV) are common pathogens that can cause acute gastroenteritis (AGE). However, very few studies have reported the molecular epidemiology and clinical information on HAstV and SaV in China. This study aims to determine the molecular epidemiology and clinical features of HAstV and SaV in patients with AGE in Guangzhou, China. Methods For this study, 656 patients with AGE were enrolled. Their stool samples were screened for 15 enteropathogens using Luminex xTAG® Gastrointestinal Pathogen Panel. HAstV and SaV were detected through an in-house multiplex reverse transcriptase polymerase chain reaction followed by phylogenetic analysis. We described and compared clinical features of AGE in patients with HAstV and SaV. Results Of the 656 stool samples, 63.72% (418/656) were found to be positive, with 550 enteropathogens (296 bacteria and 254 viruses). HAstV and SaV were detected in 20 (3.0%) and 12 (1.8%) samples, respectively. Four genotypes (genotypes 1, 2, 3, and 8) of HAstV and three genotypes (GI.1, GI.2 and GIV) of SaV were identified. Coinfection was observed in ten HAstV-positive and two SaV-positive samples. HAstV was more likely to occur in winter, while SaV in early spring. The median age of the patients with single HAstV infection was higher than that of the patients with other viruses (rotavirus, norovirus, and enteric adenovirus; P = 0.0476) and unknown etiology (P = 0.006). Coinfection with HAstV or SaV were not associated with disease severity (P > 0.05). Conclusion HAstV and SaV are the common causes of AGE in Guangzhou, China.

目的 分析急性上呼吸道感染儿童患者的病原学及临床特征.方法 以南方医科大学珠江医院2009年11月至2015年9月收治的2 665例急性上呼吸道感染儿童为研究对象,采用qRT-PCR方法检测临床上常见的8种呼吸道病毒(流感病毒、呼吸道合胞病毒、副流感病毒、腺病毒、人类博卡病毒、人类冠状病毒、人类偏肺病毒、鼻病毒).结果 共检测患儿标本2 665份,其中阳性标本1 566份,总阳性率为58.8％.四个季节中8种呼吸道病毒检出率存在明显差异,并以春季最高,夏冬季次之,秋季最低.儿童呼吸道病毒感染率随着年龄增加而逐渐降低,并以0～1岁婴幼儿病毒检出率最高64.5％.男童呼吸道病毒感染率高于女童,住院患儿呼吸道病毒检出率高于门诊患儿.混合感染标本260份,占阳性标本数的16.6％,主要集中于0～3岁儿童患者标本中,并因季节而异,秋冬季节较少,而春夏季节较为普遍.咳嗽为呼吸道病毒感染的主要临床症状,咳痰和流涕次之,临床症状在8种呼吸道病毒感染患儿中存在差异.结论 本调查分析了急性上呼吸道感染患儿中8种常见呼吸道病毒的病原学及临床特征,为指导临床治疗及防控提供相关数据.

目的 建立以白念珠菌Csa2蛋白抗体为靶标的双抗原夹心ELISA,初步评价其鉴别诊断白念珠菌尿道感染的应用价值.方法 通过间接免疫荧光染色定位Csa2蛋白在白念珠菌菌体分布情况;应用棋盘滴度法优化反应条件,建立双抗原夹心ELISA并进行方法学评价;检测324例健康体检者血清,确定该方法的cutoff值;检测120例健康体检者及30例念珠菌属尿路感染患者血清,分析其诊断的灵敏性、特异性、阳性预测值、阴性预测值.结果 Csa2蛋白是一种特异性分布在白念珠菌菌丝态表面的蛋白;以0.1μg/mL的rCsa2蛋白为包被抗原,1∶1 000稀释的rCsa2-HRP蛋白为检测抗原,建立双抗原夹心ELISA,可最低检测1∶16 000稀释的rCsa2蛋白免疫兔血清并不与正常兔血清发生反应;该方法的cutoff值为0.072,诊断白念珠菌尿路感染敏感性为65.2％ (15/23),特异性为98.4％ (125/127),阳性预测值88.2％(15/17),阴性预测值为94.0％(125/133).结论 成功建立检测白念珠菌特异性Csa2蛋白抗体双抗原夹心ELISA;初步评价该方法对诊断白念珠菌尿路感染具有较高的特异性,得出较高阳性预测值和阴性预测值;Csa2抗体靶标对区分白念珠菌感染和定植具有一定价值.

OBJECTIVES:To compare effects of repairing injured tunica albuginea (PTA) of rat penis by single or repeated local injections of chlorhexidine ethanol (ChE) into the PTA and to establish a new animal model of Peyronie's disease (PD).MATERIALS AND METHODS:Forty-two rats were divided into 7 groups. Rats either served as the normal control group with 1-5 injections of 0.9% saline or they received a single injection, 2, 3, 4, or 5 injections of ChE (0.1% chlorhexidine gluconate plus 15% ethanol dissolved in saline); rats in the positive control group were injected with TGF-β1. At 60 days after the last injection, the intracavernous pressure, degree of penile curvature, and histology were evaluated.RESULTS:Compared with the single injection of the ChE group, we found the following in the repeat ChE injections groups: an increase in the degree of penile curvature, fibrous plaques in the PTA and/or corpus cavernosum, broken elastic fibers, slightly decreased erectile function, and an increased expression of TGF-β1 and αSMA.CONCLUSIONS:Repeated ChE injuries of PTA may lead to fibrosis. This represents an excellent model of PD that involves repeated injections of ChE into the local PTA as well as reveals the pathophysiologic mechanism of PD.

The activation of tunica albuginea myofibroblasts (MFs) serves an essential role in Peyronie's disease (PD). Increasing evidence has reported that adipose tissue-derived stem cells (ADSCs) have been demonstrated to attenuate the symptoms of PD in animal models. However, the mechanisms of the antifibrotic effects of ADSCs in PD remain to be fully elucidated. In the present study, the inhibitory effects and possible mechanism of ADSCs on the activation of MFs derived from rat penile tunica albuginea were investigated. ADSCs were obtained from the paratesticular fat of Sprague Dawley rats. MFs were transformed from rat penile tunica albuginea fibroblasts through stimulation with 5 ng/ml tumor growth factor-β1. Transwell cell cultures were adopted for co-culture of ADSCs and MFs. Western blot analysis was used to assess changes in the expression levels of α smooth muscle actin (αSMA), collagen I, phosphorylated (p)-SMAD family member 2 (Smad2), Smad2, ras homolog family member A (RhoA), Rho associated coiled-coil containing protein kinase (ROCK)1 and ROCK2, caspase3, caspase9, and matrix metalloproteinases (MMPs). Collagen gel assays were used to assess cell contractility. Additionally, the concentration of hydroxyproline in the culture medium was detected using commercially available kits. It was demonstrated that ADSCs reduced the expression of αSMA and collagen I of MFs. Furthermore, p-Smad2, RhoA, ROCK1 and ROCK2 expression was significantly reduced in the MFs+ADSCs group compared with that in the MFs-only culture, while the expression of MMPs (MMP2, MMP3, MMP9 and MMP13) and caspases (caspase3 and caspase9) was upregulated. In addition, ADSCs were able to downregulate the concentration of hydroxyproline in the culture medium of MFs and reverse the contraction of MFs. Collectively, these results suggested that ADSCs inhibited the activation of MFs, decreased collagen production, and suppressed the contraction of myofibroblasts, via Smad and RhoA/ROCK signaling pathways. Furthermore, ADSCs reduced the deposition of collagen and promoted the apoptosis of MFs via MMPs, and caspases. Accordingly, the application of ADSCs may provide a novel therapeutic strategy for PD.

Objective To study the viral etiologic characteristics of acute respiratory viruses associated with adults influenza like illness (ILI) case.Methods ILI cases and non-ILI cases of nose swabs specimen were collected from Zhujiang Hospital of Southern Medical University from Nov 2009 to Oct 2014.qRT-PCR was used to test for 18 respiratory viruses.Results 201 virus positive responses were detected in samples from 420 (47.9％) patients with influenza like illness,while only 3 cases (2.6％) were found to be infected with virus in 115 non-ILI cases.The most common virus identified was influenza virus (IFV) in frequency of 51.2％ (103/201),followed by human rhinovirus (RHV) and coronaviruses (Coy).There was no significant difference among each age group in virus detection rate,however,there were obvious differences in viruses detected of several age groups,which showed that the main inflect crowd of influenza was 18 to 30 age group,the main inflect crowd of human coronavirus was 51-77 age group,and the main inflect crowd of human rhinovirus was 51-90 age group.Conclusion ILI cases in adult mosdy were caused by infection with several kinds of respiratory virus,and the dominant virus for ILI was proved to be the seasonal influenza viruses.

Understanding the molecular basis of the neutralizing antibody response to dengue virus (DENV) is an essential component in the design and development of effective vaccines and immunotherapeutics. Here we present the structure of a cross-reactive, neutralizing antibody, 3E31, in complex with domain III (DIII) of the DENV envelope (E) protein and reveal a conserved, temperature-sensitive, cryptic epitope on DIII that is not available in any of the known conformations of E on the dengue virion. We observed that 3E31 inhibits E-mediated membrane fusion, suggesting that the antibody is able to neutralize virus through binding an as-yet uncharacterized intermediate conformation of DENV E and sterically block trimer formation. Finally, we show that, unlike cross-reactive fusion peptide-specific antibodies, 3E31 does not promote antibody-dependent enhancement of infection at sub-neutralizing concentrations. Our results highlight the 3E31 epitope on the E protein DIII as a promising target for immunotherapeutics or vaccine design.

目的 以Ⅰ型登革病毒(DENV)非结构蛋白1(NS1)为免疫原制备NS1特异性单克隆抗体,建立Ⅰ型登革病毒NS1抗原特异性检测方法,并应用于广东以Ⅰ型登革感染为主的临床血清标本检测.方法 用原核表达的重组登革病毒NS1蛋白和灭活的Ⅰ型登革病毒免疫Balb/c小鼠,通过细胞融合,间接ELISA、免疫荧光和免疫印迹鉴定,获得DENV1NS1特异性单克隆抗体并建立检测方法.结果 共获得26株Ⅰ型登革病毒NS1蛋白特异性结合的单抗.通过对26株1型登革病毒NS1蛋白特异性单抗进行多株抗体组合配对,最终筛选出最佳抗体组合.该组合检测DENV1病毒培养上清的检测限为1∶16 384(12.5 pfu/ml),且与其他3个血清型登革病毒、乙型脑炎病毒和黄热病毒均无交叉反应,对500份健康人血清标本检测中,阴性标本497例,临界3例,对发病0～13d的836份临床确诊的登革热患者血清检测的敏感性为85.6％,明显高于qRT-PCR检测体系(66.3％)(P＜0.001),亦明显高于病毒分离诊断率(P＜0.001).结论 成功获得了1组DENV1-NSl蛋白的特异性单克隆抗体,利用该组抗体建立了1种敏感性和特异性更好的只针对DENV1-NS1蛋白的双抗体夹心检测方法,且比其他方法能够覆盖更宽泛的时间范围,该方法可作为登革热早期诊断的工具.

Background: Since 1997, the Asia-Pacific region had experienced epidemics of Enterovirus 71 (EV71)-associated Hand-Foot-Mouth Disease (HFMD), with cases at risk for severe illness and even death.EV71 and coxsackie virus A group 16 (CA16) are both major causative agents of HFMD and have similar early symptoms. These similarities hamper early diagnosis, making it difficult to identify potentially severe cases without genotyping. EV71-VP1 gene sequence are related to the severity of disease remains controversial. Method: We performed a retrospective study of clinical cases in Guangdong Province, China, from April 2009 to December 2012, using real-time RT-PCR to detect EV71/CA16/pan-enterovirus. Viral isolation followed VP1 genes sequencing were performed for molecular epidemiological analyses. Result: 395 and 156 hospitalized patients were confirmed to be infected with EV71 and CA16, accounting for 51.6% and 20.4% of all inpatients with enterovirus infections, respectively. Hyperpyrexia (≥39 oC, P<0.001), vomiting (P<0.001), headache (P=0.03), and neurological symptoms such as irritability (P<0.001), altered level of consciousness (P<0.001), tremors/trembling in the extremities (P<0.001), limb weakness/paralysis (P=0.007), and altered muscular tension (P=0.02) were significantly more common in EV71than in CA16-infected patients. The incidence of neurological complications (62.8% VS 5.1%, P<0.001) and outcomes were significantly different in two infections. Logistic-regression analysis revealed four independent risk factors: chloride-ion concentration, LDH activity in the CSF, troponin I levels, and serum myoglobin. EV71 isolates were all C4a subgeno-group, sharing high identity with each other (94.4%–100%). Amino acid sequences of three EV71 strains from severe cases were identical to those from mild cases. Conclusion: A comparative study of the differences in the clinical presentations of EV71 and CA16 infections in China yielded insights. Clinical features strongly associated with rapid progression and severe EV71 infections were identified. This retrospective study provides valuable information to primary-care doctors, permitting rapid intervention for high-risk patients. No difference in EV71-VP1 amino sequences were found between severe and mild cases.

The envelope domain III (EDIII) of the dengue virus (DENV) has been confirmed to be involved in receptor binding. It is the target of specific neutralizing antibodies, and is considered to be a promising subunit dengue vaccine candidate. However, several recent studies have shown that anti‑EDIII antibodies contribute little to the neutralizing or enhancing ability of human DENV‑infected serum. The present study involved an analysis of the neutralization and antibody‑dependent enhancement (ADE) activities of EDIII‑reactive antibodies in human convalescent sera from patients with primary DENV‑1 infection and rabbit antiserum immunized with recombinant DENV‑1 EDIII protein. The results indicated that serum neutralization was not associated with titres of EDIII‑binding antibodies in the human DENV‑1‑infected sera. The depletion of anti‑EDIII antibodies from these serum samples revealed that the anti‑EDIII antibodies of the patients contributed little to neutralization and ADE. However, the EDIII‑reactive antibodies from the rabbit antiserum exhibited protective abilities of neutralization at a high dilution (~1:50,000) and ADE at a low dilution (~1:5,000) for the homotypic DENV infection. Notably, the rabbit antiserum displayed ADE activity only at a dilution of 1:40 for the heterotypic virus infection, which suggests that EDIII‑reactive antibodies may be safe in secondary infection with heterotypic viruses. These results suggest that DENV EDIII is not the predominant antigen of the DENV infection process; however, purified or recombinant DENV EDIII may be used as a subunit vaccine to provoke an effective and safe antibody response.

Human adenoviruses (HAdVs) are highly contagious pathogens causing acute respiratory disease (ARD), such as community-acquired pneumonia. HAdV-7d, a re-emergent genomic variant, has been recently reported in Asia and the United States after a several-decade absence. However, whether HAdV-7d is associated with higher severity than other types is currently unclear. In this study, the clinical and epidemiological investigation showed that fever, cough, and sore throat were the three most common respiratory symptoms of HAdV infections. HAdV-7 caused longer duration of fever, higher morbidity of tachypnea/dyspnea, pleural effusion, diarrhea, hepatosplenomegaly, consciousness alteration, as well as higher rates of pneumonia, mechanical ventilation and higher fatality rate (28.6%) than other types, particularly HAdV-3 and HAdV-2. The genomes of seven HAdV-7d isolates from mild, severe, and fatal cases were sequenced and highly similar with each other. Surprisingly, two isolates (2011, 2012) had 100% identical genomes with an earlier strain from a fatal ARD outbreak in China (2009), which elucidates the virus origin and confirms the unexpected HAdV genomic conservation and stability. Phylogenetic analysis indicated that L1 52/55-kDa DNA packaging protein may be associated with the higher severity of illness and fatality rate of HAdV-7. Clinicians need to be aware of HAdVs in children with ARD.

目的 初步了解广州地区儿童和成人病毒性腹泻的病原学构成和分布情况及相关病毒的分子特征.方法 收集2012年11月-2013年5月于珠江医院就诊的腹泻患者的粪便标本,采用免疫层析法检测轮状病毒和腺病毒抗原,实时荧光PCR检测诺如病毒GⅠ/GⅡ,上述检测阳性标本进行病毒基因测序分型.结果 290例患者中,3种常见病毒的检出率分别为21.4％、15.5％和1.7％;男性患者病毒阳性率显著高于女性患者(x2=0.017,P＜0.05);≤5岁患者轮状病毒阳性率最高,为28.49％(x2=0.017,P＜0.05),＞5岁患者诺如病毒阳性率最高,为21.62％;轮状病毒性腹泻秋冬季节高发;轮状病毒G/P分型以G1P8、G9P8、G2P4和G3P8为主,占92.16％,诺如病毒共检测出8种基因型,以GⅡ.4型为主,占53.33％;病毒合并其他常见腹泻病原体感染占病毒阳性标本的15.89％.结论 轮状病毒和诺如病毒分别是广州地区婴幼儿和成人病毒性腹泻的主要病原体,本地区流行的轮状病毒和诺如病毒优势株分别是G1P8和GⅡ.4,诺如病毒基因型多样性更丰富;混合感染常见,需引起重视.

This study performed to identify and characterize the epitopes recognized by cross-neutralizing antibody against envelope protein domain Ⅲ(ED Ⅲ) of dengue virus(DENV). Two strains of cross-neutralizing monoclonal antibody(m Ab) against ED Ⅲ of dengue virus were used to screen the epitopes from phage peptide library, and the selected phage clones were tested by sandwich enzyme-linked immunosorbent assay(ELISA), and then analyzed by sequencing and bioinformatics. Furthermore, the antigenicity of the mimotope to conservative epitope within EDⅢ was identified by murine antisera immunized with positive phage clones. Phage clones binding to m Ab 2B11A35 and 2D73A7 were obtained and proved as conformational epitopes. The bioinformatics analysis showed that m Ab 2B11A35 and 2D73A7 could recognize the epitope that located in the EF loop on EDⅢ protein.The antisera of mice immunized with 3 phage clones binding to m Ab 2B11A35 were capable of binding to dengue1, 3 and 4 ED Ⅲ proteins and DENV 2 infected cells. In conclusion, we identified a new conservative and protective epitope in EDⅢ protein recognized by two strains of cross-neutralizing m Abs, which could be used for dengue virus vaccine design.

Prevalence of disseminated Penicillium marneffei infection is not known in human immunodeficiency virus (HIV)-infected patients. This retrospective study aimed to evaluate the prevalence of and risk factors for disseminated P. marneffei infection in HIV-infected patients during 2004-11 in Guangzhou, China. We tested 8131 archived HIV-infected patient serum samples for P. marneffei-specific mannoprotein (Mplp) antigen using a highly sensitive and specific ELISA that we previously established. The CD4 count of 2686 cases was determined by flow cytometry. Logistic regression was used to assess predictors of Mplp antigenaemia. The overall prevalence of disseminated penicilliosis as detected by positive serum Mplp antigen was 9.36% (761/8131), in good concordance with Platelia (TM) Aspergillus immunoassay. During 2004-11, the prevalence increased to a peak of 12.58% (158/1256) in 2010 and decreased in 2011. Penicilliosis was strongly associated with progression from HIV to AIDS (OR 4.66, 95% CI 3.94-5.51, p <0.001) and humidity (OR 1.02, 95% Cl 1.01-1.03, p 0.002). Disseminated penicilliosis occurred mainly during the rainy seasons (p <0.001). For 2686 cases with known CD4 count, logistic regression showed that CD4 count of <200 cells/mu L was a risk factor for penicilliosis (OR 2.90, 95% CI 1.10-7.66, p 0.032), especially when it was <50 cells/mu L (OR 24.26, 95% CI 1,0.63-55.36, p <0.001) during which 28.06% of patients developed disseminated penicilliosis. In conclusion, approximately 9.36% of the HIV-infected patients in our study developed disseminated penicilliosis. Rapid diagnosis may be achieved by performing serological surveillance for Mplp antigenaemia as a routine procedure for all HIV-infected patients with CD4 count of <50 cells/mu L. Clinical Microbiology and Infecticin (C) 2014 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Objective To construct a recombinant expression vector for expression of the function-al domains of dengue virus serotype 1 ( DENV1 ) envelope ( E ) protein in native soluble form. Methods The genes encoding the functional domains of DENV1-E protein (1-394 aa) were amplified with PCR and then cloned into the Psectag2B-Fc eukaryotic expression vector.The 293T cells were transfected with the recombinant vector by cationic lipid-based delivery.The cell clones expressing the fusion DENV1-E-Fc protein were screened out with 2 mg/ml of Zeocin.Immunofluorescence assay ( IFA) was performed to analyze the antigenicity and integrity of the fusion protein.The fusion proteins were purified from cell lysate with Protein-G and further identified by Western blot assay.Results The soluble form of fusion protein with a molecular weight of about 90×103 was obtained at a yield of about 25 μg per 1×107 cells.The results of IFA indicated that the fusion protein kept its integrity with right conformational epitopes.The fusion protein was successfully expressed with the advantage of good specificity as indicated by IFA and Western blot assay. Conclusion The recombinant fusion protein in soluble form was successfully expressed in eukaryotic ex-pression system, which paved the way for further investigation on the function of DENV1 E protein and its protective epitopes.

Objective To compare the detection efficiency between multiplex RT-PCR method and liquichip technology for screening the viral etiological agents of diarrhea.Methods The development of the multiplex RT-PCR method.A total of 107 feces samples from patients who suffered from diarrhea and attended to Zhujiang Hospital of Southern University from September 2013 to February 2014 were collected and tested in parallel by both multiplex RT-PCR and xTAG Gastrointestinal Pathogen Panel ( xTAG GPP) for Adenovirus, Norovirus genogroupⅠandⅡ, as well as by both multiplex RT-PCR and monoplex RT-PCR for Astrovirus and Sapovirus.To evaluate the sensitivity and specificity of multiplex RT-PCR, xTAG GPP and monoplex RT-PCR were used as reference.Kappa coefficient test was used to evaluate the consistency among the methods.The detection limit and accuracy of multiplex RT-PCR were evaluated by detection of serial dilution of positive plasmids and products sequencing for the five viral agents.Results The multiplex RT-PCR showed high consistency with xTAG GPP and monoplex RT-PCR, in which Kappa value was 0.885 and 1.000 respectively( P=0.000 ).Compared to xTAG GPP, the sensitivity and specificity of the multiplex RT-PCR were at average of 80.8%( 21/26 ) and 100%( 295/295 ) respectively.The detection limit and accuracy of multiplex RT-PCR were 104 copies /μl-106 copies/μl.Conclusion The high consistency indicated that both the multiplex RT-PCR and xTAG GPP are useful as a special,sensitive, high throughput and rapid diagnostic tools for the detection of the major viral pathogens related to diarrhea in clinical laboratory.