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Yuhua Qi,
Lunbiao Cui,
Yiyue Ge,
Zhiyang Shi,
Kangchen Zhao,
Xiling Guo,
Dandan Yang,
Hao Yu,
Lan Cui,
Yunfeng Shan,
Minghao Zhou, Hua Wang,
Zuhong Lu
[show abstract]
[hide abstract]
ABSTRACT: BACKGROUND: Pulmonary tuberculosis (TB) is a highly lethal infectious disease and early diagnosis of TB is critical for the control of disease progression. The objective of this study was to profile a panel of serum microRNAs (miRNAs) as potential biomarkers for the early diagnosis of pulmonary TB infection. METHODS: Using TaqMan Low-Density Array (TLDA) analysis followed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) validation, expression levels of miRNAs in serum samples from 30 patients with active tuberculosis and 60 patients with Bordetella pertussis (BP), varicella-zoster virus (VZV) and enterovirus (EV) were analyzed. RESULTS: The Low-Density Array data showed that 97 miRNAs were differentially expressed in pulmonary TB patient sera compared with healthy controls (90 up-regulated and 7 down-regulated). Following qRT-PCR confirmation and receiver operational curve (ROC) analysis, three miRNAs (miR-361-5p, miR-889 and miR-576-3p) were shown to distinguish TB infected patients from healthy controls and other microbial infections with moderate sensitivity and specificity (area under curve (AUC) value range, 0.711-0.848). Multiple logistic regression analysis of a combination of these three miRNAs showed an enhanced ability to discriminate between these two groups with an AUC value of 0.863. CONCLUSIONS: Our study suggests that altered levels of serum miRNAs have great potential to serve as non-invasive biomarkers for early detection of pulmonary TB infection.
BMC Infectious Diseases 12/2012; 12(1):384. · 3.12 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
[show abstract]
[hide abstract]
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
[show abstract]
[hide abstract]
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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Yiyue Ge,
Kangchen Zhao,
Yuhua Qi,
Xiaoyan Min,
Zhiyang Shi,
Xian Qi,
Yunfeng Shan,
Lan Cui,
Minghao Zhou,
Yong Wang, Hua Wang,
Lunbiao Cui
[show abstract]
[hide abstract]
ABSTRACT: Pertussis is a highly contagious, respiratory disease associated with substantial morbidity and mortality. A rapid and reliable diagnostic method is essential for appropriate treatment and prevention. Expression profiles of circulating microRNAs (miRNAs) have been proven as new non-invasive biomarkers for infectious diseases. We aimed to investigated the serum miRNA profile in pertussis patients and explored its potential as a novel diagnostic biomarker for pertussis. Among 664 different miRNAs analyzed using a miRNA array, 50 were overexpressed and 81 were underexpressed in the serum of pertussis patients. Expression levels of seven candidate miRNAs were further evaluated by real-time qRT-PCR. A panel of five miRNAs (miR-202, miR-342-5p, miR-206, miR-487b, miR-576-5p) was confirmed overexpressed in pertussis patients (p < 0.05). Risk score and receiver-operating characteristic (ROC) analysis showed that the area under the curve of the five-member miRNA profile was 0.980. At an optimal cutoff value (0.707), this panel of miRNAs yielded a sensitivity of 97.4 % and a specificity of 94.3 %. These data suggest that the five-member serum miRNA profile may serve as a new biomarker for pertussis diagnosis with high specificity and sensitivity.
Molecular Biology Reports 10/2012; · 2.93 Impact Factor
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ABSTRACT: Currently, three predominant subtypes of influenza virus are prevalent in pig populations worldwide: H1N1, H3N2, and H1N2. European avian-like H1N1 viruses, which were initially detected in European pig populations in 1979, have been circulating in pigs in eastern China since 2007. In this study, six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China. Based on whole genome sequencing, molecular characteristics of two isolates were determined. Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations, especially similar to those found in China. Four potential glycosylation sites were observed at positions 13, 26, 198, 277 in the HA1 proteins of the two isolates. Due to the presence of a stop codon at codon 12, the isolates contained truncated PB1-F2 proteins. In this study, the isolates contained 591Q, 627E and 701N in the polymerase subunit PB2, which had been shown to be determinants of virulence and host adaptation. The isolates also had a D rather than E at position 92 of the NS1, a marker of mammalian adaptation. Both isolates contained the GPKV motif at the PDZ ligand domain of the 3' end of the NS1, a characteristic marker of the European avian-like swine viruses since about 1999, which is distinct from those of avian, human and classical swine viruses. The M2 proteins of the isolates have the mutation (S31N), a characteristic marker of the European avian-like swine viruses since about 1987, which may confer resistance to amantadine and rimantadine antivirals. Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs, and raise more concerns about the occurrence of cross-species transmission events.
Virologica Sinica 10/2012; 27(5):292-8.
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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
[show abstract]
[hide abstract]
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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[hide abstract]
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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Feng-Cai Zhu,
Jun-Zhi Wang,
Xiu-Ling Li,
Zheng-Lun Liang,
Heng-Ming Ge,
Fan-Yue Meng,
Qun-Ying Mao,
Yun-Tao Zhang,
Zhen-Yu Zhang,
Hong Ji, [......],
Qing-Hua Chen,
Xiao-Qin Chen,
Wei-Wei Zhang,
Yue-Mei Hu,
Liang Li,
Feng-Xiang Li,
Xin Yao,
Pei Liu, Hua Wang,
Xin-Liang Shen
[show abstract]
[hide abstract]
ABSTRACT: : Enterovirus 71 (EV71) is highly contagious and can cause severe complications. A safe and effective vaccine is needed. We assessed the reactogenicity and immunogenicity of an inactivated, alum-adjuvanted EV71 vaccine in this study.
: A randomized, double-blind, placebo-controlled clinical trial was undertaken in 360 healthy participants who were stratified into 2 age groups (6-12 and 13-60 months), and randomly allocated to receive placebo or the investigational vaccine containing 160 U, 320 U or 640 U antigen per dose by the ratio of 1:1:1:1 at days 0 and 28. Reactogenic data within 28 days after each vaccination were recorded. Blood samples were obtained on days 0, 28 and 56 for neutralizing antibody assay.
: Overall, 193 participants reported at least 1 injection-site or systemic adverse reaction with 53.3% and 54.4% participants receiving the study vaccine and placebo, respectively. Most of the reactions were mild or moderate. Three serious adverse events were observed, but none was related to vaccination. In the participants with seronegative baseline, after 2 doses all the participants receiving EV71 vaccines were seropositive and the seroconversion rates were more than 98.1%. In the participants with seropositive baseline, 1 dose induced good seroconversion rates of more than 64.3% in participants receiving EV71 vaccines.
: This study found that the inactivated EV71 vaccine was well tolerated and had good immunogenicity in healthy children and infants. A single dose induced typical booster response in the participants with a seropositive baseline, and 2 doses were needed for the immunologically naive participants.
The Pediatric Infectious Disease Journal 08/2012; 31(11):1158-65. · 3.58 Impact Factor
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[hide abstract]
ABSTRACT: Borehole acoustic reflection logging can provide high resolution images of near-borehole geological structure. However, the
conventional seismic migration and imaging methods are not effective because the reflected waves are interfered with the dominant
borehole-guided modes and there are only eight receiving channels per shot available for stacking. In this paper, we apply
an equivalent offset migration method based on wave scattering theory to process the acoustic reflection imaging log data
from both numerical modeling and recorded field data. The result shows that, compared with the routine post-stack depth migration
method, the equivalent offset migration method results in higher stack fold and is more effective for near-borehole structural
imaging with low SNR acoustic reflection log data.
Applied Geophysics 04/2012; 6(4):303-310. · 0.45 Impact Factor
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Yanwen Xiong,
Ping Wang,
Ruiting Lan,
Changyun Ye, Hua Wang,
Jun Ren,
Huaiqi Jing,
Yiting Wang,
Zhemin Zhou,
Xuemei Bai,
Zhigang Cui,
Xia Luo,
Ailan Zhao,
Yan Wang,
Shaomin Zhang,
Hui Sun,
Lei Wang,
Jianguo Xu
[show abstract]
[hide abstract]
ABSTRACT: An Escherichia coli O157:H7 outbreak in China in 1999 caused 177 deaths due to hemolytic uremic syndrome. Sixteen outbreak associated isolates were found to belong to a new clone, sequence type 96 (ST96), based on multilocus sequence typing of 15 housekeeping genes. Whole genome sequencing of an outbreak isolate, Xuzhou21, showed that the isolate is phylogenetically closely related to the Japan 1996 outbreak isolate Sakai, both of which share the most recent common ancestor with the US outbreak isolate EDL933. The levels of IL-6 and IL-8 of peripheral blood mononuclear cells induced by Xuzhou21 and Sakai were significantly higher than that induced by EDL933. Xuzhou21 also induced a significantly higher level of IL-8 than Sakai while both induced similar levels of IL-6. The expression level of Shiga toxin 2 in Xuzhou21 induced by mitomycin C was 68.6 times of that under non-inducing conditions, twice of that induced in Sakai (32.7 times) and 15 times higher than that induced in EDL933 (4.5 times). Our study shows that ST96 is a novel clone and provided significant new insights into the evolution of virulence of E. coli O157:H7.
PLoS ONE 01/2012; 7(4):e36144. · 4.09 Impact Factor
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Feng-Cai Zhu,
Zheng-Lun Liang,
Fan-Yue Meng,
Ying Zeng,
Qun-Ying Mao,
Kai Chu,
Xue-Fang Song,
Xin Yao,
Jing-Xin Li,
Hong Ji,
Yi-Ju Zhang,
Liang Li,
Hong-Xing Pan,
Ke Xu,
Wei-Ming Dai,
Wei-Wei Zhang,
Fei Deng, Hua Wang,
Jun-Zhi Wang
[show abstract]
[hide abstract]
ABSTRACT: Hand, foot, and mouth disease (HFMD) has been emerging as an important public problem over the past few decades, especially in Asian and Pacific regions. A national program on EV71 vaccine development against HFMD was initiated in China, in 2008, which called for a need for seroepidemiological study for the target population.
This was a retrospective study conducted in Jiangsu Province, in October, 2010. We measured the neutralizing antibodies against EV71 and CoxA16 in a cohort of infants aged of 2, 7, 12, and 27-38 months and their mothers just before delivery. Series sera samples from 975 infants and 555 mothers were collected and analyzed. Questionnaires on the history of HFMD were completed in the survey. A total of 143 HFMD cases were collected, but only 11.2% were reported to the National Infectious Disease Information Management System. The level of maternal antibody titers decreased dramatically during the first 7 month and remained at a relatively low level thereafter. But it increased significantly from month 12 to months 27-38. The accumulate incidence density of HFMD demonstrated a significant increase after 14 months of age, resulting in a accumulate incidence density of 50.8/1000 person-years in survey period. Seropositivity of EV71 antibody in infants at the age of 2 months seems to demonstrate a protective effect against HFMD.
High seropositive rate of EV71 and CoxA16 antibody was found in prenatal women in mainland China, and there is a need to enhance the HFMD case management and the current surveillance system. We suggest that infants aged between 6 to 14 months should have the first priority to receive EV71 vaccine.
PLoS ONE 01/2012; 7(5):e37206. · 4.09 Impact Factor
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Chang-jun Bao,
Xi-ling Guo,
Xian Qi,
Jian-li Hu,
Ming-hao Zhou,
Jay K Varma,
Lun-biao Cui,
Hai-tao Yang,
Yong-jun Jiao,
John D Klena, [......],
Yin Chen,
Zheng Zhu,
Ke Xu,
Ai-hua Shen,
Tao Wu,
Hai-yan Peng,
Zhi-feng Li,
Jun Shan,
Zhi-yang Shi, Hua Wang
[show abstract]
[hide abstract]
ABSTRACT: Seven persons in one family living in eastern China developed fever and thrombocytopenia during May 2007, but the initial investigation failed to identify an infectious etiology. In December 2009, a novel bunyavirus (designated severe fever with thrombocytopenia syndrome bunyavirus [SFTSV]) was identified as the cause of illness in patients with similar clinical manifestations in China. We reexamined this family cluster for SFTSV infection.
We analyzed epidemiological and clinical data for the index patient and 6 secondary patients. We tested stored blood specimens from the 6 secondary patients using real time reverse transcription polymerase chain reaction (RT-PCR), viral culture, genetic sequencing, micro-neutralization assay (MNA), and indirect immunofluorescence assay (IFA).
An 80-year-old woman with fever, leucopenia, and thrombocytopenia died on 27 April 2007. Between 3 and 7 May 2007, another 6 patients from her family were admitted to a local county hospital with fever and other similar symptoms. Serum specimens collected in 2007 from these 6 patients were positive for SFTS viral RNA through RT-PCR and for antibody to SFTSV through MNA and IFA. SFTSV was isolated from 1 preserved serum specimen. The only shared characteristic between secondary patients was personal contact with the index patient; none reported exposure to suspected animals or vectors.
Clinical and laboratory evidence confirmed that the patients of fever and thrombocytopenia occurring in a family cluster in eastern China in 2007 were caused by a newly recognized bunyavirus, SFTSV. Epidemiological investigation strongly suggests that infection of secondary patients was transmitted to family members by personal contact.
Clinical Infectious Diseases 12/2011; 53(12):1208-14. · 9.15 Impact Factor
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Yongjun Jiao,
Xiaoyan Zeng,
Xiling Guo,
Xian Qi,
Xiao Zhang,
Zhiyang Shi,
Minghao Zhou,
Changjun Bao,
Wenshuai Zhang,
Yan Xu, Hua Wang
[show abstract]
[hide abstract]
ABSTRACT: The recent emergence of the human infection confirmed to be caused by severe fever with thrombocytopenia syndrome virus (SFTSV) in China is of global concern. Safe diagnostic immunoreagents for determination of human and animal seroprevalence in epidemiological investigations are urgently needed. This paper describes the cloning and expression of the nucleocapsid (N) protein of SFTSV. An N-protein-based double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) system was set up to detect the total antibodies in human and animal sera. We reasoned that as the double-antigen sandwich ELISA detected total antibodies with a higher sensitivity than traditional indirect ELISA, it could be used to detect SFTSV-specific antibodies from different animal species. The serum neutralization test was used to validate the performance of this ELISA system. All human and animal sera that tested positive in the neutralization test were also positive in the sandwich ELISA, and there was a high correlation between serum neutralizing titers and ELISA readings. Cross-reactivity was evaluated, and the system was found to be highly specific to SFTSV; all hantavirus- and dengue virus-confirmed patient samples were negative. SFTSV-confirmed human and animal sera from both Anhui and Hubei Provinces in China reacted with N protein in this ELISA, suggesting no major antigenic variation between geographically disparate virus isolates and the suitability of this assay in nationwide application. ELISA results showed that 3.6% of the human serum samples and 47.7% of the animal field serum samples were positive for SFTSV antibodies, indicating that SFTSV has circulated widely in China. This assay, which is simple to operate, poses no biohazard risk, does not require sophisticated equipment, and can be used in disease surveillance programs, particularly in the screening of large numbers of samples from various animal species.
Journal of clinical microbiology 11/2011; 50(2):372-7. · 4.16 Impact Factor
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New England Journal of Medicine 09/2011; 365(9):862-3; author reply 864-5. · 53.30 Impact Factor
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Xue-Jie Yu,
Mi-Fang Liang,
Shou-Yin Zhang,
Yan Liu,
Jian-Dong Li,
Yu-Lan Sun,
Lihong Zhang,
Quan-Fu Zhang,
Vsevolod L Popov,
Chuan Li, [......],
Xiu-Ping Fu,
Li-Na Sun,
Xiao-Ping Dong,
Zi-Jian Feng,
Wei-Zhong Yang,
Tao Hong,
Yu Zhang,
David H Walker,
Yu Wang,
De-Xin Li
[show abstract]
[hide abstract]
ABSTRACT: Heightened surveillance of acute febrile illness in China since 2009 has led to the identification of a severe fever with thrombocytopenia syndrome (SFTS) with an unknown cause. Infection with Anaplasma phagocytophilum has been suggested as a cause, but the pathogen has not been detected in most patients on laboratory testing.
We obtained blood samples from patients with the case definition of SFTS in six provinces in China. The blood samples were used to isolate the causal pathogen by inoculation of cell culture and for detection of viral RNA on polymerase-chain-reaction assay. The pathogen was characterized on electron microscopy and nucleic acid sequencing. We used enzyme-linked immunosorbent assay, indirect immunofluorescence assay, and neutralization testing to analyze the level of virus-specific antibody in patients' serum samples.
We isolated a novel virus, designated SFTS bunyavirus, from patients who presented with fever, thrombocytopenia, leukocytopenia, and multiorgan dysfunction. RNA sequence analysis revealed that the virus was a newly identified member of the genus phlebovirus in the Bunyaviridae family. Electron-microscopical examination revealed virions with the morphologic characteristics of a bunyavirus. The presence of the virus was confirmed in 171 patients with SFTS from six provinces by detection of viral RNA, specific antibodies to the virus in blood, or both. Serologic assays showed a virus-specific immune response in all 35 pairs of serum samples collected from patients during the acute and convalescent phases of the illness.
A novel phlebovirus was identified in patients with a life-threatening illness associated with fever and thrombocytopenia in China. (Funded by the China Mega-Project for Infectious Diseases and others.).
New England Journal of Medicine 03/2011; 364(16):1523-32. · 53.30 Impact Factor
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ABSTRACT: Usually, magnetic nanoparticles (MNPs) are prepared based on the famous Stöber process in which divinylbenzene (DVA) is often used as a crosslink agent to synthesize SiO2/(PMMA/Fe3O4) nanoparticles. Compared with DVA, linolenic acid (LNA) is innoxious and can polymerize more easily for it has three unsaturated double bonds. In this paper, LNA was used as a new crosslink agent instead of DVA to synthesize the SiO2/(PMMA/Fe3O4) nanoparticles. The results showed that the core-shell structure could be observed obviously. The sizes of nanoparticles with core-shell structure range from 200 to 500 nm. The DNA probes which was immobilized on the surface of MNPs were used to capture the biotin modified complementary sequence of the probe, and the formed complexes were bonded with streptavidin-modified alkaline phosphatase (SA-AP). Finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy) phenyl-1, 2-dioxetane (AMPPD) which was the substrate reagent of AP. The specificity and sensitivity of this approach were investigated in this paper.
Journal of Nanoscience and Nanotechnology 03/2011; 11(3):2256-62. · 1.56 Impact Factor
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[show abstract]
[hide abstract]
ABSTRACT: A novel method was established through the detection of chemiluminescent signals of nucleic acid hybridization based on magnetic nanoparticles (MNPs) and PCR. 5' amino- modified specific probes were immobilized on the surface of silanized MNPs by Schiff reaction between amino and aldehyde group. The probes were used to capture the synthetic biotin-dUTP-labeled DNA fragments which were obtained by polymerase chain reaction (PCR). Then these complexes were bonded with streptavidin-modified alkaline phosphatase (SA-AP). Finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)- 4-methoxy -4-(3"-phosphoryloxy) phenyl-1, 2-dioxetane (AMPPD) which was the substrate reagent of AP. The concentration of probes which were immobilized on the surface of MNPs was studied, how to reduce the adsorption of SA-AP on the surface of MNPs was also researched. It was shown that 12.5 pmol of probes were immobilized on 1 mg of MNPs. Aldehyde-MNPs modified with probes could adsorb SA-AP, affecting the sensitivity of chemiluminescene consequently. Reduction of aldehyde group by sodium borohydride and blocking the bare position of MNPs with bovine serum albumin (BSA) could decrease the background of chemiluminescence, and this method has good specificity in detection of chloramphenicol acetyltransferase (CAT) gene.
Journal of Nanoscience and Nanotechnology 02/2011; 11(2):1074-8. · 1.56 Impact Factor
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Ping Wang,
Yanwen Xiong,
Ruiting Lan,
Changyun Ye, Hua Wang,
Jun Ren,
Huaiqi Jing,
Yiting Wang,
Zhemin Zhou,
Zhigang Cui, [......],
Dong Jin,
Xuemei Bai,
Ailan Zhao,
Yan Wang,
Shaomin Zhang,
Hui Sun,
Juan Li,
Tao Wang,
Lei Wang,
Jianguo Xu
[show abstract]
[hide abstract]
ABSTRACT: In addition to the large virulence plasmid pO157, a novel 38-kb conjugative plasmid, pO157_Sal, was identified and sequenced from an Escherichia coli O157:H7 outbreak-associated Chinese isolate that shares high similarity with a plasmid in Salmonella enterica serovar Agona. The plasmid was found in 15 of 326 isolates, 12 of which were of the same pulsed-field gel electrophoresis type.
Journal of clinical microbiology 02/2011; 49(4):1594-7. · 4.16 Impact Factor
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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
[show abstract]
[hide abstract]
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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Lunbiao Cui,
Yuhua Qi,
Haijing Li,
Yiyue Ge,
Kangchen Zhao,
Xian Qi,
Xiling Guo,
Zhiyang Shi,
Minghao Zhou,
Baoli Zhu,
Yan Guo,
Jun Li,
Charles W Stratton,
Yi-Wei Tang, Hua Wang
[show abstract]
[hide abstract]
ABSTRACT: Altered circulating microRNA (miRNA) profiles have been noted in patients with microbial infections. We compared host serum miRNA levels in patients with hand-foot-and-mouth disease (HFMD) caused by enterovirus 71 (EV71) and coxsackievirus 16 (CVA16) as well as in other microbial infections and in healthy individuals. Among 664 different miRNAs analyzed using a miRNA array, 102 were up-regulated and 26 were down-regulated in sera of patients with enteroviral infections. Expression levels of ten candidate miRNAs were further evaluated by quantitative real-time PCR assays. A receiver operating characteristic (ROC) curve analysis revealed that six miRNAs (miR-148a, miR-143, miR-324-3p, miR-628-3p, miR-140-5p, and miR-362-3p) were able to discriminate patients with enterovirus infections from healthy controls with area under curve (AUC) values ranged from 0.828 to 0.934. The combined six miRNA using multiple logistic regression analysis provided not only a sensitivity of 97.1% and a specificity of 92.7% but also a unique profile that differentiated enterovirial infections from other microbial infections. Expression levels of five miRNAs (miR-148a, miR-143, miR-324-3p, miR-545, and miR-140-5p) were significantly increased in patients with CVA16 versus those with EV71 (p<0.05). Combination of miR-545, miR-324-3p, and miR-143 possessed a moderate ability to discrimination between CVA16 and EV71 with an AUC value of 0.761. These data indicate that sera from patients with different subtypes of enteroviral infection express unique miRNA profiles. Serum miRNA expression profiles may provide supplemental biomarkers for diagnosing and subtyping enteroviral HFMD infections.
PLoS ONE 01/2011; 6(11):e27071. · 4.09 Impact Factor
