期刊文献+
共找到793篇文章
< 1 2 40 >
每页显示 20 50 100
Detection of hepatitis B virus DNA by real-time PCR using TaqMan-MGB probe technology 被引量:15
1
作者 Jin-RongZhao Yu-JieBai +3 位作者 Qing-HuaZhang YanWan DingLi Xiao-JunYan 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第4期508-510,共3页
AIM: To develop a real-time PCR for detecting hepatitis B virus-(HBV) DNA based on TaqMan technology using a new MGB probe.METHODS: Plasmid containing the sequence of X gene (1414-1744 nt) was constructed as HBV-DNA s... AIM: To develop a real-time PCR for detecting hepatitis B virus-(HBV) DNA based on TaqMan technology using a new MGB probe.METHODS: Plasmid containing the sequence of X gene (1414-1744 nt) was constructed as HBV-DNA standard for quantitative analysis. A TaqMan-MGB probe between primers for amplification was designed to detect PCR products. The interested sequence contained in the plasmid and in clinical specimens was quantitatively measured.RESULTS: The detection limit of the assay for HBV DNA was 1 genome equivalent per reaction. A linear standard curve was obtained between 100 and 109 DNA copies/reaction (r>0.990). None of the negative control samples showed false-positive reactions in duplicate. HBV DNA was detected in 100% (50/50) of HBV patients with HbeAg, and in 72.0% (36/50) with HBsAg, HBeAb and HBcAb. The coefficient of variation for both intra- and inter-experimental variability demonstrated high reproducibility and accuracy.CONCLUSION: Real-time PCR based on TaqMan-MGB probe technology is an excellent method for detection of HBV DNA. 展开更多
关键词 hepatitis B virus DNA TaqMan-MGB probe Real-time pcr
下载PDF
Rapid quantification of hepatitis B virus DNA by real-time PCR using efficient TaqMan probe and extraction of virus DNA 被引量:9
2
作者 Yan-Qin Lu Jin-Xiang Han +3 位作者 Peng Qi Wei Xu Yan-Hui Zu Bo Zhu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第45期7365-7370,共6页
AIM: To rapidly quantify hepatitis B virus (HBV) DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. METHODS: Three standards were prepared by cloning PCR products which targeted... AIM: To rapidly quantify hepatitis B virus (HBV) DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. METHODS: Three standards were prepared by cloning PCR products which targeted S, C and X region of HBV genome into pGEM-T vector respectively. A pair of primers and matched TaqMan probe were selected by comparing the copy number and the Ct values of HBV serum samples derived from the three different standard curves using certain serum DNA. Then the efficiency of six HBV DNA extraction methods including guanidinium isothiocyanate, proteinase K, NaI, NaOH lysis, alkaline lysis and simple boiling was analyzed in sample A, B and C by real-time PCR. Meanwhile, 8 clinical HBV serum samples were quantified. RESULTS: The copy number of the same HBV serum sample originated from the standard curve of S, C and X regions was 5.7 × 10^4/mL, 6.3 × 10^2/mL and 1.6 × 10^3/ mL respectively. The relative Ct value was 26.6, 31.8 and 29.5 respectively. Therefore, primers and matched probe from S region were chosen for further optimization of six extraction methods. The copy number of HBV serum samples A, B and C was 3.49 × 10^9/mL, 2.08 × 10^6/mL and 4.40 × 10^7/mL respectively, the relative Ct value was 19.9, 30 and 26.2 in the method of NaOH lysis, which was the efficientest among six methods. Simple boiling showed a slightly lower efficiency than NaOH lysis. Guanidinium isothiocyanate, proteinase K and NaI displayed that the copy number of HBV serum sample A, B and C was around 10^S/mL, meanwhile the Ct value was about 30. Alkaline failed to quantify the copy number of three HBV serum samples. Standard deviation (SD) and coefficient variation (CV) were very low in all 8 clinical HBV serum samples, showing that quantification of HBV DNA in triplicate was reliable and accurate. CONCLUSION: Real-time PCR based on optimized primers and TaqMan probe from S region in combination with NaOH lysis is a simple, rapid and accurate method for quantification of HBV serum DNA. 展开更多
关键词 hepatitis B virus Serum DNA Real-time pcr Extraction method
下载PDF
Oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus 被引量:3
3
作者 Yong-Zhong Wang Guo-Xiang Wu Li-Bo Luo Min Chen Li-Hua Ruan 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第31期4260-4263,共4页
To compare the oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus in Chinese patients with chronic hepatitis B. METHODS: Mixture of samples with different genotypes and clinical se... To compare the oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus in Chinese patients with chronic hepatitis B. METHODS: Mixture of samples with different genotypes and clinical serum samples from 126 chronic hepatitis B patients was tested for hepatitis B virus genotypes by oligonucleotide chip, real-time PCR and sequencing of PCR products, respectively. Clinical performances, time required and costs of the three assays were evaluated. RESULTS: Oligonucleotide chips and real-time PCR detected 1% and 0.1% genotypes, respectively, in mixed samples. Of the 126 clinical samples from patients with chronic hepatitis B, genotype B was detected in 41 (33%), 41 (33%) and 45 (36%) samples, and genotype C in 76 (60%), 76 (60%) and 81 (64%) samples, by oligonucleotide chip, real-time PCR and sequencing, respectively. Oligonucleotide chip and real-time PCR detected mixed genotypes B and C in 9 samples. Real- time PCR was the rapidest and cheapest among the three assays. CONCLUSION: Oligonucleotide chip and real-time PCR are able to detect mixed genotypes, while sequencing only detects the dominant genotype in clinical samples. 展开更多
关键词 hepatitis B virus GENOTYPES Oligonucleotidechips Real-time pcr SEQUENCING
下载PDF
Hepatitis B virus in cerebrospinal fluid of a patient with purulent bacterial meningitis detected by multiplex-PCR:A case report 被引量:2
4
作者 Dai-Quan Gao Yong-Qiang Hu +1 位作者 Xin Wang Yun-Zhou Zhang 《World Journal of Clinical Cases》 SCIE 2022年第5期1697-1701,共5页
BACKGROUND Bacterial meningitis(BM)is a common central nervous system inflammatory disease.BM may cause serious complications,and early diagnosis is essential to improve the prognosis of affected patients.CASE SUMMARY... BACKGROUND Bacterial meningitis(BM)is a common central nervous system inflammatory disease.BM may cause serious complications,and early diagnosis is essential to improve the prognosis of affected patients.CASE SUMMARY A 37-year-old man was hospitalized with purulent meningitis because of worsening headache for 12 h,accompanied by vomiting,fever,and rhinorrhea.Head computed tomography showed a lesion in the left frontal lobe.Infectious disease screening showed positivity for hepatitis B surface antigen,hepatitis B e antigen,and hepatitis B core antigen.Cerebrospinal fluid(CSF)leak was suspected based on clinical history.Streptococcus pneumoniae(S.pneumoniae)was detected in CSF by metagenomic next-generation sequencing(mNGS)technology,confirming the diagnosis of purulent BM.After treatment,multiplex PCR indicated the presence of hepatitis B virus(HBV)DNA and absence of S.pneumoniae DNA in CSF samples.CONCLUSION We report a rare case of HBV in the CSF of a patient with purulent BM.Multiplex PCR is more sensitive than mNGS for detecting HBV DNA. 展开更多
关键词 Purulent meningitis Streptococcus pneumoniae hepatitis B virus Multiplex pcr Cerebrospinal fluid Case report
下载PDF
Determining hepatitis C virus genotype distribution among high-risk groups in Iran using real-time PCR 被引量:1
5
作者 Marzieh Jamalidoust Mandana Namayandeh +2 位作者 Sadaf Asaei Nasrin Aliabadi Mazyar Ziyaeyan 《World Journal of Gastroenterology》 SCIE CAS 2014年第19期5897-5902,共6页
AIM: To assess hepatitis C virus (HCV) genotype patterns among high-risk Iranian groups, using real-time RT-PCR.
关键词 hepatitis C virus genotype distribution Injection drug users Real-time pcr Iran
下载PDF
Molecular Characterization and SYBR Green I-Based Quantitative PCR for Duck Hepatitis Virus Type 1 被引量:1
6
作者 LUO Yu-jun ZHANG Gui-hong +2 位作者 XU Xiao-qin CHEN Jian-hong LIAO Ming 《Agricultural Sciences in China》 CAS CSCD 2008年第9期1140-1146,共7页
To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain, real-time quantitative polymerase chain reaction (RTQ-PCR) assay based on SYBR Green I technology was developed to target 3D gene ... To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain, real-time quantitative polymerase chain reaction (RTQ-PCR) assay based on SYBR Green I technology was developed to target 3D gene of DHV-1, Comparative sequence analysis showed that the genome has a typical picornarivus genetic organization, and strain DHV-1 R genetic organaiztion is 5' untranslated region (UTR)-VP0-VP3-VPI-2A1-2A2-2B-2C-3A-3B-3C-3D-3' UTR, DHV-1 R has close relationship with Parechovirus, and has 95.1-99.1% nucleotide sequence identity with other DHV-1 strains. Based on the DHV-1 sequences in GenBank, three pairs of specific primers were designed to amplify DHV-1 using real-time PCR. The results showed that real-time PCR Tm value is 85.6℃ and the real-time PCR provides a broad dynamic range, detecting from 102 to 109 copies of DHV-1 cDNA per reaction. No cross-reactions were found in specimens containing DPV, AIV and NDV. It is concluded that DHV-1 belongs to a new group of the family Picornaviridae that may form a separate genus most closely related to the genus Parechovirus. All results showed that the real-time PCR has high sensitivity and specificity to detect DHV-1 using SYBR Green I dissociation curve analysis, isolates can be distinguished by their melting temperature. These methods are rapid, sensitive, and reliable, and can be readily adapted for detection of DHV-1 from other clinical samples. 展开更多
关键词 duck hepatitis virus type 1 complete genome real-time RT-pcr
下载PDF
Development of RT PCR Assay for Detection of Duck Hepatitis Virus Type 1
7
作者 Wang Yongjuan Cui Pingfu +1 位作者 Liu Bo Meng Ting 《Animal Husbandry and Feed Science》 CAS 2017年第5期308-310,共3页
In this study,a RT-PCR assay for rapid detection of duck hepatitis virus type 1( DHV-1) was established in the presence of a pair of primers designed based on vp1 gene sequence,using the genomic RNA of DHV-1 or other ... In this study,a RT-PCR assay for rapid detection of duck hepatitis virus type 1( DHV-1) was established in the presence of a pair of primers designed based on vp1 gene sequence,using the genomic RNA of DHV-1 or other common viruses as the template. The template was serially diluted 10-fold to determine the reaction sensitivity. Finally,seven liver samples of ducks with suspected DHV-1 infection,which were collected from different regions of Jiangsu Province,were detected with this method. The results showed that a DNA fragment of about 360 bp was specifically amplified with the RT-PCR system,and the detection limit was1 fg/μL. The detection rate of clinical samples with this method was 100%. The results revealed that the RT-PCR system which had high specificity and high sensitivity in the detection of DHV-1 was successfully established. 展开更多
关键词 DUCK hepatitis virus TYPE 1 RT pcr DETECTION
下载PDF
Hepatitis A Virus PCR Analysis and <i>E. coli</i>Detection in Oysters at Oualidia Lagoon and Their Correlation
8
作者 Naima El Moqri Najwa Hassou +3 位作者 Fatiha El Mellouli Hasnae Zekhnini Nabil Abouchoaib Samira Etahiri 《Food and Nutrition Sciences》 2020年第7期684-694,共11页
The present study aims to evaluate hepatitis A virus (HAV) prevalence and faecal contamination indicators <span style="font-family:Verdana;"><i></i></span><i><i><span s... The present study aims to evaluate hepatitis A virus (HAV) prevalence and faecal contamination indicators <span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">Escherichia coli</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;"> (</span><span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">E. coli</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;">) in oysters from Oualidia lagoon (Moroccan Atlantic coast) and to study the correlation between the two parameters. The survey was carried out on 87 samples of oysters (</span><span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">Crassostrea gigas</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;">) collected monthly between November 2015 and February 2017 from three sites corresponding to different oyster farms in the lagoon. Sanitary status of bivalve molluscs was assessed by </span><span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">E. coli</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;"> enumeration using ISO 16649-3. Detection of hepatitis A virus, was carried out by real-time reverse transcriptase polymerase chain reaction (rRT-PCR) according to ISO 15216-2 method. The prevalence of samples for which </span><span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">E. coli</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;"> contamination exceeds the threshold of 230 </span><span style="font-family:Verdana;"><i></i></span><i><i><span style="font-family:Verdana;">E. coli</span></i><i><span style="font-family:Verdana;"></span></i></i><span style="font-family:Verdana;">/100g of flesh and intravalvular fluid (FIF) is 43%. HAV RNA was detected in 2% of the samples analyzed. This RNA was even detected in a sample meeting the bacterial criteria. Viral health surveillance of bivalve molluscs is therefore necessary before their delivery for human consumption.</span> 展开更多
关键词 hepatitis A virus E. coli Bivalves SHELLFISH RT-pcr Oualidia Lagoon
下载PDF
Detection on hepatitis c virus of blood samples with fluorescence quantitative PCR
9
《中国输血杂志》 CAS CSCD 2001年第S1期405-,共1页
关键词 Detection on hepatitis c virus of blood samples with fluorescence quantitative pcr
下载PDF
Detection of the covalently closed circular DNA of duck hepatitis B virus by Taq-Man fluorescent quantitative PCR assay
10
作者 MEI LI FU QING LIN +3 位作者 XIAO PENG LIU SHUI LAN SHI DONG LIANG LI ZI RONG CHEN 《Journal of Microbiology and Immunology》 2007年第1期35-39,共5页
To develop a fluorescent quantitative PCR assay based on Taq-Man chemistry to detect the covalenfly closed circular DNA (eccDNA) of duck hepatitis B virus (DHBV), a pair of primers was designed from both sides of ... To develop a fluorescent quantitative PCR assay based on Taq-Man chemistry to detect the covalenfly closed circular DNA (eccDNA) of duck hepatitis B virus (DHBV), a pair of primers was designed from both sides of the nick in the minus strand of DHBV and a Taq-Man probes between the primers, modified with 6-Fam at 5' end and Tamra at its 3' end was designed to detect the PCR products during PCR cycles. The DHBV DNA fragment was cloned into vector PUCm-T, and the recombinant plasmid was purified and subsequently qualified as the HBV DNA standard. The experimental conditions and reagents used in PCR assay for amplification were sophisticatedly optimized in order to yield a perfect amplification efficacy and reduce the possibility to produce non-specific amplification. It was demonstrated that the detect limit of assay was 10^3 copies/ml, and a linear standard curve was obtained between 10^5 -10^9 copies/ml [ C1 =-2.8361 ln(x) + 41.45, r =-0.9985]. The coefficient of variation was 0.2%-3.14% and 2.22%-4.43% for intra- and inter-assay respectively. After a dynamic survey on the contents of DHBV DNA in serum of ducks, it was found that its peak value appeared at the second week of birth in ducks. It is evident that this method of Taq-Man fluorescent quantitative PCR assay appears to be simple, sensitive and specific. 展开更多
关键词 DUCK hepatitis B virus Covalently closed circular DNA(cccDNA) Fluorescence quantitative pcr
下载PDF
A sensitive method to detect the hepatitis B virus mutations by using solid phase PCR on oligonucleotide array
11
作者 YAN QIN LU JIN XIANG HAN +1 位作者 ZHONG LIN SHEN CHUAN XI WANG 《Journal of Microbiology and Immunology》 2006年第4期265-271,共7页
A sensitive method based on solid phase PCR on oligonucleotide array was established to detect two point mutations 1896G-A and 1901G-A in hepatitis B virus (HBV) DNA, in which 6 probes including these two point mutati... A sensitive method based on solid phase PCR on oligonucleotide array was established to detect two point mutations 1896G-A and 1901G-A in hepatitis B virus (HBV) DNA, in which 6 probes including these two point mutations were immobilized on modified glass slides through 5' terminal linker, while the 3' terminal was made to be free. The mutated loci were designed to locate on the last nucleotides of 3' terminal respectively, and the positive control probes lacked the last nucleotide of 3' terminal in comparison with the probes used for detection. Probes fixed on oligonucleotide array were also the solid phase amplification primers. One pair of liquid primers was used to amplify the short template product from whole HBV DNA. Using target DNA as template, the solid primers were extended under the action of Taq DNA polymerase and incorporated with Cy-3dCTP as marker. During the thermal cycling reaction, probes served as solid phase amplification primers and amplification products bound to the oligonucleotide array, which could be visualized by incorporation with fluorescent dyes. After amplification, the oligonucleotide array was washed, performed with laser scanning, and then used for quantitative analysis to obtain the information for mutations. The hybridization results were compared with DNA sequencing. It was demonstrated that in case of sample A, the ratios of fluorescence intensities in wide type and in the mutated types of 1896G-A and 1901G-A mutations in HBV were 3.81:1 and 1:3.79 respectively, while, in case of sample B, those were 1:2.89 and 1:3.03 respectively, indicating the presence of point mutations in these two loci. These results correlated to those obtained from DNA sequencing analysis in which the fluorescence intensity ratios in wide type and in the mutated types of 1996G-A and 1901D-A mutations in HBV were 1.26:1 and 1.67:1 respectively. From the above observations, it is evident that the method using solid phase PCR based on oligonucleotide array appears to be a sensitive and promising way to detect mutations with low-density. 展开更多
关键词 Oligonucleotide array Solid phase pcr hepatitis B virus Point mutation
下载PDF
Analysis of hepatitis B virus preS1 variability and prevalence of the rs2296651 polymorphism in a Spanish population 被引量:5
12
作者 Rosario Casillas David Tabernero +9 位作者 Josep Gregori Irene Belmonte Maria Francesca Cortese Carolina González Mar Riveiro-Barciela Rosa Maria López Josep Quer Rafael Esteban Maria Buti Francisco Rodríguez-Frías 《World Journal of Gastroenterology》 SCIE CAS 2018年第6期680-692,共13页
AIM To determine the variability/conservation of the domain of hepatitis B virus(HBV) pre S1 region that interacts with sodium-taurocholate cotransporting polypeptide(hereafter, NTCP-interacting domain) and the preval... AIM To determine the variability/conservation of the domain of hepatitis B virus(HBV) pre S1 region that interacts with sodium-taurocholate cotransporting polypeptide(hereafter, NTCP-interacting domain) and the prevalence of the rs2296651 polymorphism(S267 F, NTCP variant) in a Spanish population. METHODS Serum samples from 246 individuals were included and divided into 3 groups: patients with chronic HBV infection(CHB)(n = 41, 73% Caucasians), patients with resolved HBV infection(n = 100, 100% Caucasians) and an HBV-uninfected control group(n = 105, 100% Caucasians). Variability/conservation of the amino acid(aa) sequences of the NTCPinteracting domain,(aa 2-48 in viral genotype D) and a highly conserved pre S1 domain associated with virion morphogenesis(aa 92-103 in viral genotype D) were analyzed by next-generation sequencing and compared in 18 CHB patients with viremia > 4 log IU/mL. The rs2296651 polymorphism was determined in all individuals in all 3 groups using an in-house real-time PCR melting curve analysis.RESULTS The HBV pre S1 NTCP-interacting domain showed a high degree of conservation among the examined viral genomes especially between aa 9 and 21(in the genotype D consensus sequence). As compared with the virion morphogenesis domain, the NTCPinteracting domain had a smaller proportion of HBV genotype-unrelated changes comprising > 1% of the quasispecies(25.5% vs 31.8%), but a larger proportion of genotype-associated viral polymorphisms(34% vs 27.3%), according to consensus sequences from Gen Bank patterns of HBV genotypes A to H. Variation/conservation in both domains depended on viral genotype, with genotype C being the most highly conserved and genotype E the most variable(limited finding, only 2 genotype E included). Of note, proline residues were highly conserved in both domains, and serine residues showed changes only to threonine or tyrosine in the virion morphogenesis domain. The rs2296651 polymorphism was not detected in any participant.CONCLUSION In our CHB population, the NTCP-interacting domain was highly conserved, particularly the proline residues and essential amino acids related with the NTCP interaction, and the prevalence of rs2296651 was low/null. 展开更多
关键词 hepatitis B virus hepatitis B virus PRES1 region sodium-taurocholate co-transporting polypeptide NTCP-interacting DOMAIN VIRION morphogenesis DOMAIN SNP rs2296651 next-generation sequencing real-time pcr melting curves
下载PDF
Comparison of real-time polymerase chain reaction with the COBAS Amplicor test for quantitation of hepatitis B virus DNA in serum samples 被引量:7
13
作者 Ming Shi Yong Zhang +2 位作者 Ying-Hua Zhu Jing Zhang Wei-Jia Xu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第3期479-483,共5页
AIM:To compare the clinical performance of a real-time PCR assay with the COBAS Amplicor Hepatitis B Virus (HBV) Monitor test for quantitation of HBV DNA in serum samples. METHODS: The reference sera of the Chinese Na... AIM:To compare the clinical performance of a real-time PCR assay with the COBAS Amplicor Hepatitis B Virus (HBV) Monitor test for quantitation of HBV DNA in serum samples. METHODS: The reference sera of the Chinese National Institute for the Control of Pharmaceutical and Biological Products and the National Center for Clinical Laboratories of China, and 158 clinical serum samples were used in this study. The linearity, accuracy, reproducibility, assay time, and costs of the real-time PCR were evaluated and compared with those of the Cobas Amplicor test. RESULTS: The intra-assay and inter-assay variations of the real-time PCR ranged from 0.3% to 3.8% and 1.4% to 8.1%, respectively. The HBV DNA levels measured by the real-time PCR correlated very well with those obtained with the COBAS Amplicor test (r = 0.948). The real-time PCR HBV DNA kit was much cheaper and had a wider dynamic range. CONCLUSION: The real-time PCR assay is an excellent tool for monitoring of HBV DNA levels in patients with chronic hepatitis B. 展开更多
关键词 COBAS Amplicor test hepatitis B virus Viral DNA Real-time pcr
下载PDF
Performance verification and comparison of TianLong automatic hypersensitive hepatitis B virus DNA quantification system with Roche CAP/CTM system 被引量:1
14
作者 Ming Li Lin Chen +9 位作者 Li-Ming Liu Yong-Li Li Bo-An Li Bo Li Yuan-Li Mao Li-Fang Xia Tong Wang Ya-Nan Liu Zheng Li Tong-Sheng Guo 《World Journal of Gastroenterology》 SCIE CAS 2017年第37期6845-6853,共9页
AIM To investigate and compare the analytical and clinical performance of Tian Long automatic hypersensitive hepatitis B virus(HBV) DNA quantification system and Roche CAP/CTM system.METHODS Two hundred blood samples ... AIM To investigate and compare the analytical and clinical performance of Tian Long automatic hypersensitive hepatitis B virus(HBV) DNA quantification system and Roche CAP/CTM system.METHODS Two hundred blood samples for HBV DNA testing, HBV-DNA negative samples and high-titer HBV-DNA mixture samples were collected and prepared. National standard materials for serum HBV and a worldwide HBV DNA panel were employed for performance verification. The analytical performance, such as limit of detection, limit of quantification, accuracy, precision, reproducibility, linearity, genotype coverage and cross-contamination, was determined using the Tian Long automatic hypersensitive HBV DNA quantification system(TL system). Correlation and Bland-Altman plot analyses were carried out to compare the clinical performance of the TL system assay and the CAP/CTM system. RESULTS The detection limit of the TL system was 10 IU/m L, and its limit of quantification was 30 IU/m L. The differences between the expected and tested concentrations of the national standards were less than ± 0.4 Log10 IU/m L, which showed high accuracy of the system. Results of the precision, reproducibility and linearity tests showed that the multiple test coefficient of variation(CV) of the same sample was less than 5% for 102-106 IU/m L; and for 30-108 IU/m L, the linear correlation coefficient r2 = 0.99. The TL system detected HBV DNA(A-H) genotypes and there was no cross-contamination during the "checkerboard" test. When compared with the CAP/CTM assay, the two assays showed 100% consistency in both negative and positive sample results(15 negative samples and 185 positive samples). No statistical differences between the two assays in the HBV DNA quantification values were observed(P > 0.05). Correlation analysis indicated a significant correlation between the two assays, r2 = 0.9774. The Bland-Altman plot analysis showed that 98.9% of the positive data were within the 95% acceptable range, and the maximum difference was-0.49.CONCLUSION The TL system has good analytical performance, and exhibits good agreement with the CAP/CTM system in clinical performance. 展开更多
关键词 Analytical performance hepatitis B virus DNA quantification Clinical performance hepatitis B virus Real-time quantification pcr
下载PDF
Antiviral effect of Chinese medicine jiaweisinisan in hepatitis B virus transgenic mice 被引量:1
15
作者 Xiao-Yin Chen Guang-Dong Tong Fang Xia 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第14期2280-2283,共4页
AIM: To study the antiviral effect of Chinese medicine jiaweisinisan (JWSNS) on hepatitis B virus (HBV) infection in transgenic mice (TGM). METHODS: Twenty two 6-8 wk old HBV TGM in the third generation were d... AIM: To study the antiviral effect of Chinese medicine jiaweisinisan (JWSNS) on hepatitis B virus (HBV) infection in transgenic mice (TGM). METHODS: Twenty two 6-8 wk old HBV TGM in the third generation were divided into TGM control group and TGM treated group randomly. The normal control group included ten normal BC 57L/6 mice at the same age. The mice in treated group were administrated with JWSNS at the concentration of 4 g/mL and the dosage of 50 g/kg per d for 30 d, while the mice in TGM control group and normal control group were administrated with normal saline at the same dosage and the same time. Polymerase chain reaction (PCR) was used to assess the contents of HBV DNA in serum of HBV TGM before and after treatments, whereas blot hybridization was utilized to measure the contents of HBV DNA in the liver of both HBV TGM and normal BC 57L/6 mice. RESULTS: The levels of serum HBV DNA in TGM treated group were remarkably decreased after the treatment of JWSNS (7.662±0.78 vs 5.22±3.14, P〈0.05), while there was no obvious change after administration of normal saline in TGM control group (7.125±4.26 vs 8.932 ± 5.12, P〉 0.05). The OD values of HBV DNA in the livers of the mice in TGM treated group were significantly lower than those of TGM control group (0.274±0.096 vs 0.432 ± 0.119, P 〈 0.01). CONCLUSION: JWSNS exerts suppressive effects on HBV DNA in the serum and liver of TGM. 展开更多
关键词 Traditional Chinese herbs hepatitis B virus Transgenic mice pcr Blot hybridization Chronic hepatitis B
下载PDF
Hepatitis B virus genotypes in chronic liver disease patients from New Delhi,India 被引量:1
16
作者 Saket Chattopadhyay Bhudev Chandra Das Premashis Kar 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第41期6702-6706,共5页
AIM: To study the Hepatitis B virus (HBV) genotypes and their effect on the progression and outcome in patients with chronic liver diseases from New Delhi, India. METHODS: Sera from 100 HBV-related chronic liver disea... AIM: To study the Hepatitis B virus (HBV) genotypes and their effect on the progression and outcome in patients with chronic liver diseases from New Delhi, India. METHODS: Sera from 100 HBV-related chronic liver disease (CLDB) cases were tested for HBV genotype using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Type-specific primers-based PCR (TSP-PCR) targeting to the surface (S) gene encoding hepatitis B surface antigen. RESULTS: Only genotypes A and D were present and genotype D was dominant. Genotype D was present in all CLDB patient categories. The genotype distribution for the 100 patients with CLDB was as follows: genotype A, 16/100 (16%) (7/40- 17% chronic hepatitis B (CHB); 8/47, 17%, HBV-related cirrhosis (CRB); 1/13, 7.6%, HBV-related hepatocellular carcinoma (HCCB); genotype D- 84/100 (84%) (32/40- 80% CHB; 38/47- 81%, CRB; 11/13, 85%, HCCB); genotype A + D, 3/100 (3%) (1/40- 3% CHB; 1/47- 2%, CRB; 1/13, 7.6%, HCCB); C, 0; B, 0; E, 0; F, 0; G 0, H 0; (P < 0.01, genotype D vs A). CONCLUSION: Only HBV genotypes A and D were present in patients with CLDB from New Delhi, India. Compared with genotype D, genotype A patients had no significant clinical or biochemical differences (P > 0.05). Mixed infection with genotype A and D were seen in 3% of the cases. Genotype D was the dominant genotype prevalent in all patient categories. 展开更多
关键词 HBV-related chronic liver disease hepatitis B virus genotypes pcr-RFLP Type-specific primer-based pcr
下载PDF
Detection of hepatitis D virus by cDNA microarray method
17
《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2004年第3期423-427,共5页
关键词 hepatitis D virus pcr MICROARRAY HYBRIDIZATION
下载PDF
Prevalence and clinical significance of SEN virus infection in patients with non A-E hepatitis and volunteer blood donors in Shanghai
18
作者 Zheng-Hao Tang Xiao-Hua Chen Yong-Sheng Yu Guo-Qing Zang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第26期4204-4208,共5页
AIM:To explore the prevalence of SEN virus (SENV) in patients with non A-E hepatitis and volunteer blood donors in Shanghai. METHODS: According to the published gene sequences, primers from the conserved region were d... AIM:To explore the prevalence of SEN virus (SENV) in patients with non A-E hepatitis and volunteer blood donors in Shanghai. METHODS: According to the published gene sequences, primers from the conserved region were designed. Then, the prevalence of SEN virus in 30 samples from healthy voluntary blood donors and 30 samples from patients with non A-E hepatitis were detected by nested-PCR of SENV-D/H. Some PCR products were cloned and sequenced. RESULTS: The specificity of genotype-specific PCR was confirmed by sequencing, the SENV DNA was detected in 53.3% of the patients with non A-E hepatitis and 10% of the blood donors. The prevalence of SENV-D/H viremia was significantly higher in patients with non A-E hepatitis than in blood donors (P = 0.0002). SENV-H subtype and SENV-D subtype were found in 2 and 1 samples, respectively from blood donors. SENV-H subtype, SENV D subtype, mixed SENV-D and SENV-H subtype were found in 8, 6 and 2 samples, respectively, from patients with non A-E hepatitis. CONCLUSION: The gene type of SENV in patients with non A-E hepatitis and blood donors in shanghai is D or H subtype, and transfusion is not the only transmitting form of SENV. 展开更多
关键词 SEN virus Nested pcr Blood donor NonA-E hepatitis
下载PDF
THE INVESTIGATION OF DIFFERENT POPULATION'S TRANSFUSION-TRANSMITTED VIRUS-DNA IN SHAANXI PROVINCE
19
作者 党双锁 程延安 +1 位作者 曾嵘 程训明 《Academic Journal of Xi'an Jiaotong University》 2000年第2期129-131,共3页
Objective To investigate transfusion transmitted virus (TTV) infection among population of different groups in Shaanxi Province.Methods A nested polymerase chain reaction (PCR) with primers from ORF1 of TTV genome was... Objective To investigate transfusion transmitted virus (TTV) infection among population of different groups in Shaanxi Province.Methods A nested polymerase chain reaction (PCR) with primers from ORF1 of TTV genome was established to detect TTV DNA in serum of the patients.Results TTV DAN was detected in the sera of 3 of 50 cases of general population(6%), 2 of 30 cases of vocational blood donors(6.7%),21 of 97 cases with Type B hepatitis(21.6%),9 of 35 cases of Type C hepatitis (25.7%),and 23 of 40 cases with non A^non G hepatitis(57 5%).Conclusion There is TTV infection among general population in Shaanxi Province.TTV may be an important agent to cause non A^non G hepatitis .And the patients with HBV or HCV can have overlapping TTV infection. 展开更多
关键词 transfusion transmitted virus(TTV) polymerase chain reaction(pcr) non-A-non-G hepatitis
下载PDF
猪肉及制品中HEV、PEDV和PDCoV三重qPCR方法的建立与应用 被引量:1
20
作者 余姓鸿 张婧 +7 位作者 安微 杨苗 谢礼 舒佳新 薛昌华 郑巧 林华 韩国全 《食品工业科技》 CAS 北大核心 2024年第2期210-219,共10页
人兽共患病引发的动物源性食品安全事件频发,为从食品原料源头上控制和保障其食用安全,控制猪肉及其制品消费成本,本研究构建了可同时检测猪肉及制品中戊型肝炎病毒(Hepatitis E virus,HEV)、猪流行性腹泻病毒(Porcine epidemic diarrhe... 人兽共患病引发的动物源性食品安全事件频发,为从食品原料源头上控制和保障其食用安全,控制猪肉及其制品消费成本,本研究构建了可同时检测猪肉及制品中戊型肝炎病毒(Hepatitis E virus,HEV)、猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)和猪δ冠状病毒(Porcine delta corona virus,PDCoV)三重实时荧光定量PCR(Real-time Quantitative PCR,qPCR)方法。结果表明,三重qPCR方法只能扩增出3种目的病毒的特异性基因片段,特异性好;对HEV、PEDV和PDCoV三种病毒的最低检测限分别为6.02、6.98、6.92 copies/μL;组内和组间变异系数(CV%)在0.10%~3.00%之间,重复性好。将所建立方法应用于248份出口猪肉及制品和282份生猪粪拭子的检测,同时以相应病毒标准检测方法进行平行检测,结果显示猪肉及制品中三种病毒的检出率均为0%,与标准方法检测结果一致。生猪粪拭子中,该方法对PEDV、PDCoV和HEV三种病毒的检出率分别为1.06%、3.19%、0.35%,标准方法检出率分别为1.06%、3.19%、0%。研究表明,建立的三重qPCR检测方法能准确快速地检测猪肉及制品或生猪样品中三种病毒,为保障生鲜猪肉及其制品市场流通和阻断病毒食源性传播提供技术支持。 展开更多
关键词 猪肉 戊型肝炎病毒(HEV) 猪流行性腹泻病毒(PEDV) 猪δ冠状病毒(PDCoV) 三重qpcr
下载PDF
上一页 1 2 40 下一页 到第
使用帮助 返回顶部