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Zhifeng Li,
Xian Qi,
Minghao Zhou,
Changjun Bao,
Jianli Hu, Bin Wu,
Shenjiao Wang,
Zhongmin Tan,
Jianguang Fu,
Jun Shan,
Yefei Zhu,
Fenyang Tang
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ABSTRACT: The aim of this study was to develop and evaluate a two-tube multiplex real-time RT-PCR assay for the detection and identification of four viral hemorrhagic fever (VHF) pathogens, severe fever with thrombocytopenia syndrome virus (SFTSV), Hantaan virus (HTNV), Seoul virus (SEOV), and dengue virus (DENV), from human clinical samples. The two-tube multiplex real-time RT-PCR assay we developed has a sensitivity of 10 copies/μL for each of the targets, and the performance was linear within the range of at least 10(7) transcript copies. Moreover, we evaluated the specificity of the assay using other virus RNA as template, and found no cross-reactivity. This new assay is able to detect SFTSV, HTNV, SEOV and DENV in two reactions and brings a cost of 40 % compared to separate reactions. Evaluation of this assay with clinical serum samples from laboratory-confirmed patients and healthy donors showed 100 % clinical diagnostic sensitivity and over 99 % specificity. The assay was applied for scanning 346 clinical samples collected from patients admitted to the hospital with suspected VHF and compared with virus isolation and immunofluorescence assay (IFA). The assay indentified 59 SFTSV-, 12 HTNV-, 11 SEOV- and 9 DENV-positive samples and showed higher sensitivity. This assay thus provides a reliable and cost-effective screening tool for early clinical diagnosis of SFTSV, HTNV, SEOV and DENV in the acute phase.
Archives of Virology 03/2013; · 2.11 Impact Factor
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Zhifeng Li,
Lunbiao Cui,
Minghao Zhou,
Xian Qi,
Changjun Bao,
Jianli Hu,
Jun Shan, Bin Wu,
Shenjiao Wang,
Xiling Guo,
Yongjun Jiao,
Fenyang Tang,
Hua Wang
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ABSTRACT: A highly sensitive one-step real-time RT-PCR method using a minor-groove-binding (MGB) probe was developed for detection and quantitation of severe febrile with thrombocytopenia syndrome virus (SFTSV). The assay could discriminate SFTSV infection from other related viral diseases in human with a minimum detection limit of 10 viral RNA copies/µl and was 1,000 times more sensitive than the conventional PCR. Strong linear correlations (r(2) > 0.99) between the C(t) values and viral RNA standards over a linear range were obtained. The coefficients of variation of intra- and inter-assay reproducibility were both less than 2%. The RT-PCR was also shown to be highly specific, as no positive signals were detected for other related viruses. Evaluation of this assay with serum samples from laboratory confirmed cases and healthy donors showed 100% clinical diagnostic sensitivity and over 99% specificity. Clinical application with samples from 287 patients admitted to the hospital with suspected SFTSV infection showed that 15% were infected by SFTSV. This assay was rapid, requiring just over 2 hr, including the nucleic acid extraction step. J. Med. Virol. © 2012 Wiley Periodicals, Inc.
Journal of Medical Virology 12/2012; · 2.82 Impact Factor
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Hong Ji,
Liang Li,
Yanming Liu,
Hengming Ge,
Xushan Wang,
Jianli Hu, Bin Wu,
Jianguang Fu,
Zhenyu Zhang,
Xiaoqin Chen,
Minglei Zhang,
Qiang Ding,
Wen Bo Xu,
Fenyang Tang,
Minghao Zhou,
Hua Wang,
Fengcai Zhu
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ABSTRACT: BACKGROUND: The major etiology of hand, foot and mouth disease (HFMD) is infection with human enterovirus A (HEV-A). Among subtypes of HEV-A, coxsackievirusA16 (CoxA16) and enterovirus 71 (EV71) are major causes for recurrent HFMD among infants and children in Jiangsu Province, mainland China. Here, we analyzed maternal antibodies between prenatal women and their neonates, to determine age-specific seroprevalence of human EV71 and CoxA16 infections in infants and children aged 0 to 15 years. The results may facilitate the development of immunization against HFMD. METHODS: This study used cross-section of 40 pairs of pregnant women and neonates and 800 subjects aged 1 month to 15 years old. Micro-dose cytopathogenic effects measured neutralizing antibodies against EV71 and CoxA16. Chi-square test compared seroprevalence rates between age groups and McNemar test, paired-Samples t-test and independent-samples t-test analyzed differences of geometric mean titers. RESULTS: A strong correlation between titers of neutralizing antibody against EV71 and CoxA16 in prenatal women and neonates was observed (rEV71 = 0.67, rCoxA16 = 0.56, respectively, p < 0.05). Seroprevalence rates of anti-EV71 antibody gradually decreased with age between 0 to 6 months old, remained low between 7 to 11 months (5.0--10.0%), and increased between 1 and 4 years (22.5--87.5%). Age-specific seroprevalence rates of anti-EV71 antibody stabilized in >80% of children between 5 to 15 years of age. However, seroprevalence rates of anti-CoxA16 antibody were very low (0.0--13.0%) between 0 to 6 months of age, gradually increased between 7 months to 4 years (15.0--70.0%), and stabilized at 54.0% (108/200) between 5 to 15 years. Seroprevalence rates against EV71 and CoxA16 were low under 1 year (0.0--10.0%), and showed an age dependent increase with high seroprevalence (52.5--62.5%) between 4 and10 years of age. CONCLUSIONS: Concomitant infection of EV71 and CoxA16 was common in Jiangsu Province. Therefore, development of bivalent vaccine against both EV71 and CoxA16 is critical. The optimal schedule for vaccination may be 4 to11 months of age.
Virology Journal 10/2012; 9(1):248. · 2.34 Impact Factor
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Lunbiao Cui,
Yiyue Ge,
Xian Qi,
Gaolian Xu,
Haijing Li,
Kangchen Zhao, Bin Wu,
Zhiyang Shi,
Xiling Guo,
Lin Hu,
Qimin You,
Li-Hong Zhang,
Alexander N Freiberg,
Xuejie Yu,
Hua Wang,
Minghao Zhou,
Yi-Wei Tang
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ABSTRACT: A virus known as the severe fever with thrombocytopenia syndrome virus (SFTSV) was recently identified as the etiological agent of severe fever with thrombocytopenia syndrome (SFTS) in China. Reliable laboratory detection and identification of this virus is likely to become clinically and epidemiologically desirable. We developed a near instrument-free, simple molecular method which incorporates a reverse-transcription cross-priming amplification (RT-CPA) coupled with a vertical flow (VF) visualization strip, for rapid detection of SFTSV. The RT-CPA-VF assay targets a conserved region of the M segment of the SFTSV genome and has a limit of detection of 100 copies per reaction with no cross-reaction with other vector-borne bunyaviruses and bacterial pathogens. The performance of the RT-CPA-VF assay was determined on 175 human plasma specimens collected from 89 clinically suspected SFTS patients and 86 healthy donors. The sensitivity and specificity of the assay was 94.1% and 100.0%, respectively, when compared with a combination of virus culture and real-time RT-PCR. The entire procedure from specimen processing to result reporting can be completed within two hours. The simplicity and the near instrument-free platform make the RT-CPA-VF assay practical for point-of-care testing.
Journal of clinical microbiology 09/2012; · 4.16 Impact Factor
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Xian Qi,
Lunbiao Cui,
Yongjun Jiao,
Yuning Pan,
Xihan Li,
Rongqiang Zu,
Xiang Huo, Bin Wu,
Fengyang Tang,
Yongchun Song,
Minghao Zhou,
Hua Wang,
Carol J Cardona,
Zheng Xing
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ABSTRACT: Cross-species transmission of influenza A viruses from swine to human occurs occasionally. In 2011, an influenza A H1N1 virus, A/Jiangsu/ALS1/2011 (JS/ALS1/2011), was isolated from a boy who suffered from severe pneumonia in China. The virus is closely related antigenically and genetically to avian-like swine H1N1 viruses that have recently been circulating in pigs in China and that were initially detected in European pig populations in 1979. The isolation of JS/ALS1/2011 provides additional evidence that swine influenza viruses can occasionally infect humans and emphasizes the importance of reinforcing influenza virus surveillance in both pigs and humans.
Archives of Virology 08/2012; · 2.11 Impact Factor
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Hong Ji,
Liang Li, Bin Wu,
Ke Xu,
Xiang Huo,
Jun Shan,
Wen-dong Liu,
Wei-wei Zhang,
Fen-yang Tang,
Feng-cai Zhu,
Rong-qiang Zu
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ABSTRACT: To analyze the etiological and epidemiological characteristics of hand-foot-and-mouth disease (HFMD) seen in Jiangsu province from 2008 to 2010, and provide evidence for its prevention and control.
Based on the requirement of supervision program of HFMD, surveillance and report were done according to National Disease Supervision Information Management System. Descriptive epidemiological method, performed between 2008 and 2010, was used to analyze the time, region and population distribution and results of etiologic analysis of HFMD. Nucleic acid of enterovirus (EV) genome was detected by real-time RT-PCR.
The average incidence rate of HFMD was 86.70 per million between 2008 and 2010, the peak incidence occurred in April to July. There were significant differences among the incidence in different districts (P<0.05), and the highest incidence was seen in the densely inhabited southern areas of Jiangsu province. Most of the cases were infants and children aged less than 5 years. The number of male cases (2008: 17,008, 2009: 48 768, 2010: 50,231) was much larger than that of the female cases 2008: 9662, 2009: 29 151, 2010: 30,655. The HFMD cases with mild symptoms were caused mainly by enterovirus 71 (EV71) and coxsackievirus A16 (Cox A16) and there was difference among different years. The severe HFMD cases and deaths were mainly caused by EV71 infection.
The epidemiologic characteristics of HFMD in Jiangsu province from 2008 to 2010 had close relationship with season, population and region. The mild cases of HFMD were mainly infected with EV71 and Cox A16. However, EV71 illness seemed to be more severe and had significantly greater frequency of serious complications and fatality than the illness caused by Cox A16.
Zhonghua er ke za zhi. Chinese journal of pediatrics 04/2012; 50(4):261-6.
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Xiang Huo,
Xian Qi,
Fenyang Tang,
Rongqiang Zu,
Liang Li, Bin Wu,
Yuanfang Qin,
Hong Ji,
Jianguang Fu,
Shenjiao Wang,
Hua Tian,
Zhibin Hu,
Haitao Yang,
Minghao Zhou,
Hua Wang,
Fengcai Zhu
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ABSTRACT: We investigated the seropositive rates and persistence of antibody against pandemic (H1N1) 2009 virus (pH1N1) in pregnant women and voluntary blood donors after the second wave of the pandemic in Nanjing, China.
Serum samples of unvaccinated pregnant women (n = 720) and voluntary blood donors (n = 320) were collected after the second wave of 2009 pandemic in Nanjing. All samples were tested against pH1N1 strain (A/California/7/2009) with hemagglutination inhibition assay. A significant decline in seropositive rates, from above 50% to about 20%, was observed in pregnant women and voluntary blood donors fifteen weeks after the second wave of the pandemic. A quarter of the samples were tested against a seasonal H1N1 strain (A/Brisbane/59/2007). The antibody titers against pH1N1 strain were found to correlate positively with those against seasonal H1N1 strain. The correlation was modest but statistically significant.
The high seropositive rates in both pregnant women and voluntary blood donors suggested that the pH1N1 virus had widely spread in these two populations. Immunity derived from natural infection seemed not to be persistent well.
PLoS ONE 01/2011; 6(3):e17995. · 4.09 Impact Factor
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Fen-yang Tang,
Rong-qiang Zu,
Liang Li,
Xian Qi,
Hong Ji, Bin Wu,
Ke Xu,
Jian-guang Fu,
Yuan-fang Qin,
Fei Deng,
Hai-tao Yang
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ABSTRACT: To understand the antibody levels against pandemic influenza A (H1N1) virus (2009 H1N1) among aged ≥ 3 years population in 2009, from Jiangsu province, and to describe the distribution of 2009 H1N1.
Serum specimens were collected from natural populations at four different periods in Jiangsu, and tested with hemagglutination-inhibition (HI) assays. Rates of protective antibody against 2009 H1N1 and Geometric mean titers (GMTs) were estimated.
The rates of protective antibody against 2009 H1N1 rose with the progress of epidemics in Jiangsu, which were 3.46%, 7.59%, 16.94%, respectively in July, August and November, 2009. There were no significant differences on the rates of protective antibody between males and females at four different cross-sectional periods (P > 0.05), and no significant differences on GMTs were observed at different periods except for November 2009. Significant differences on rates of protective antibody and GMTs among various age groups were observed at four different periods (P < 0.05), and similar results were observed among different periods in various age groups (P < 0.05). There were significant differences on rates of protective antibody and GMTs among different areas (P < 0.05).
The 2009 H1N1 strain had been widely spread out in Jiangsu province since July 2009. People aged 12 - 17 years became the major victims after August. As of November 2009, the rate of protective antibody against 2009 H1N1 was still low, predicting that the epidemic might continue to exist for a certain period of time.
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 05/2010; 31(5):489-93.
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ABSTRACT: Pathogens of the first influenza pandemic this century belong to influenza A H1N1 viruses, which are different from human seasonal H1N1 viruses in antigenic and genetic characterization. To better understand the genetic characteristics and evolution, timely detect variant strains with epidemiological importance, we analyzed in detail the molecular characterization of the early influenza A H1N1 (2009) virus.
Genomic sequences of reference influenza viruses were obtained from Influenza Resource Center of GenBank. Sequences were analyzed using the EditSeq and Megalign program with the Lasergene sequence analysis software package (DNAStar, Madison, WI, USA). A/California/07/2009 (H1N1) was selected as a representative strains of the novel influenza A H1N1 (2009) virus, and its molecular characteristics was determined.
A/California/07/2009 do not contained the molecular characteristics of highly pathogenic influenza virus, and its 11 proteins retained most of the molecular characteristics of swine influenza virus, but also had some characteristics of avian and human influenza viruses. With a classical swine H1N1 and human H1N1 dual character, PB1-F2 protein of A/California/07/2009 terminates after 11aa, 57aa and 87aa, which is a unique molecular characteristics of influenza H1N1 (2009) virus.
This is the first report for detailed analysis of Molecular characteristics of the novel influenza A H1N1 (2009) virus. As the virus further adapt and persist in human populations, its molecular characteristics will change accordingly. So we should pay special attention to the effect on virus transmission and pathogenesis.
ACTA MICROBIOLOGICA SINICA 01/2010; 50(1):81-90.
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ABSTRACT: To determine the epidemiological features of hand-foot-and-mouth disease (HFMD) outbreaks and the genetic characteristics of enterovirus type 71(EV71) isolates from patients in Lianyungang, Jiangsu province in May, 2008.
Epidemiological, microbiological, cellular and molecular methods were performed to investigate pathogens and to describe the homogeneity of isolated strains.
21 cases were reported in this HFMD outbreak with the attack rate as 20.0%. 3 EV71 virus strains were isolated from 10 stool samples. The nucleotide and amino acid homogeneity of these 3 Jiangsu strain with Anhui Fuyang strains were 97.9%-100.0% and 99.7%-100.0%, respectively. These 4 Jiangsu strains were within genotype C sub-geno group C4 in phylogenetic tree. Data from the follow-up study showed that shedding of EV71 and Coxsackie A16 virus (CA16) in the latent period appeared in the outbreak of HFMD. Human beings could be infected by both EV71 virus and CA16 at the same time and could also carry the two viruses. We also discovered that EV71 virus could be expelled out of the human body through stool in the first week and last for 10 weeks.
The recently identified EV71 isolates from this HFMD outbreak belonged to sub-geno group C4. Facts as: the release of viruses in the latent period, co-infection or coexisting of two viruses at the same time and super long period of expulsion of toxin exist in EV71 and CA16 did exist.
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 05/2009; 30(4):339-43.
