1.Detection of Epstein-Barr virus DNA in plasma/serum: a useful serological indicator for diagnosis of nasopharyngeal carcinoma.
Shijuan MAI ; Yongsheng ZONG ; Min ZHANG ; Biling ZHONG ; Suxia LIN
Chinese Medical Journal 2002;115(12):1895-1897
OBJECTIVETo compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.
METHODSThe plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted.
RESULTSEBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion >/= 1:40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA >/= 1:40 test (0.4839, 0.5385 and 1.0480, respectively).
CONCLUSIONThe EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.
Antigens, Viral ; blood ; DNA, Viral ; blood ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Serologic Tests
2.Study on etiology of clinically diagnosed non A-E hepatitis.
Zhongping HE ; Hui ZHUANG ; Shujing SONG ; Xueping WANG ; Rongbing WANG ; Zhuang LIU
Chinese Journal of Experimental and Clinical Virology 2002;16(1):7-10
BACKGROUNDTo study etiology of clinically diagnosed non A-E hepatitis.
METHODSHBV, TTV, human parvovirus B19, SENV DNA were detected by nested polymerase chain reactions (nPCR), while HGV, HCV RNA were tested by reverse transcription nested polymerase chain reactions (RT-nPCR).
RESULTSOf 60 patients with clinically diagnosed non A-E hepatitis, 30 (50.0%) were HBV DNA positive alone, 10 (16.7%) HBV and TTV DNA positive, 6 (10.0%) HBV and B19 DNA positive; 1 (1.7%) HBV, SENV DNA and HCV RNA positive, 1 (1.7%) HCV RNA positive alone, 1 (1.7%) HCV RNA and B19 DNA positive, 2 (3.3%) B19 DNA positive alone, 1 (1.7%) TTV DNA positive alone, and the remaining 8 (13.3%) negative for all viruses. All the 60 patients were HGV RNA negative. There were no differences in serum biochemical markers of hepatitis B patients with or without TTV or B19 virus infection.
CONCLUSIONSHBV is a major etiologic agent for the clinically diagnosed non A-E hepatitis. HGV, TTV, B19 and SEBV may not be associated with nonA-E hepatitis.
Adult ; Aged ; DNA, Viral ; blood ; Female ; Hepacivirus ; genetics ; isolation & purification ; Hepatitis B ; diagnosis ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis, Viral, Human ; diagnosis ; virology ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; Sequence Analysis, DNA
3.Real-time fluorescent quantitative PCR assay for measuring cytomegalovirus DNA load in patients after haematopoietic stem cell transplantation.
Jun FAN ; Wei-hang MA ; Mei-fang YANG ; Han XUE ; Hai-nü GAO ; Lan-juan LI
Chinese Medical Journal 2006;119(10):871-874
Adolescent
;
Adult
;
Antigens, Viral
;
blood
;
Cytomegalovirus
;
isolation & purification
;
DNA, Viral
;
analysis
;
Female
;
Fluorescence
;
Hematopoietic Stem Cell Transplantation
;
Humans
;
Male
;
Middle Aged
;
Phosphoproteins
;
blood
;
Polymerase Chain Reaction
;
methods
;
Viral Load
;
Viral Matrix Proteins
;
blood
4.The progress of congenital cytomegalovirus infection and detection methods.
Xiao-Zhou HE ; Xiao-Fang WANG ; Shi-Wen WANG
Chinese Journal of Virology 2012;28(1):73-77
Cytomegalovirus infection is extremely common in the population, especially for newborns. Congenital CMV infection may cause central nervous system damage and other related diseases, thus potentially harmful. At home and abroad, some related research had been carried out on the incidence of disease, and a variety of detection methods had been developed. In this paper, the current situation of congenital cytomegalovirus infection and detection method is reviewed.
Antibodies, Viral
;
blood
;
Cytomegalovirus
;
isolation & purification
;
Cytomegalovirus Infections
;
congenital
;
diagnosis
;
DNA, Viral
;
blood
;
Female
;
Humans
;
Infant, Newborn
;
Polymerase Chain Reaction
;
Pregnancy
5.Distribution of HCV genotypes in volunteer blood donors in Guangzhou.
Han-shen YE ; Ru XU ; Min WANG ; Xia RONG ; Wen-feng HUA ; Xin YE ; Guang-ping RUO ; Yong-shui FU
Journal of Southern Medical University 2010;30(12):2655-2657
OBJECTIVETo determine the distribution of HCV genotypes among volunteer blood donors in Guangzhou.
METHODSSix-nine HCV RNA-positive samples were collected from volunteer blood donors in Guangzhou. NS5B fragments of HCV were amplified followed by DNA sequencing and phylogenetic analysis.
RESULTSHCV genotypes were determined for 67 samples. Among them, the subtypes 1b, 2a, 3a, 3b, 6a and 6n were detected at the frequencies of 37.31%, 4.48%, 7.46%, 4.48%, 44.78% and 1.49%, respectively.
CONCLUSIONHCV 1b and 6a are the most predominant two subtypes among volunteer blood donors in Guangzhou.
Blood Donors ; China ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, DNA
7.On the detection of HBV cccDNA and its clinical significance: an overview of research advancement.
Journal of Biomedical Engineering 2009;26(3):662-666
Covalently closed circular DNA (cccDNA) is the existing form of the HBV DNA in the nucleus of host cells and also the original template of HBV replication; its long-term presence in the nucleus makes it difficult to be eliminated by current antiviral drugs; and it becomes the key factor of continuous HBV infection and relapse after antiviral suspension. Detection of HBV cccDNA is of great significance for further understanding the life cycle of HBV and providing guidance for antiviral treatment. This article aims to review the detection and its clinical significance to the advancement of researches on hepatitis B virus cccDNA.
DNA, Circular
;
blood
;
genetics
;
DNA, Viral
;
blood
;
genetics
;
Hepatitis B virus
;
genetics
;
isolation & purification
;
Hepatitis B, Chronic
;
virology
;
Humans
;
Polymerase Chain Reaction
;
methods
8.Performance Evaluation of Abbott RealTime HBV Quantification Kit for HBV Viral Load by Real-Time PCR.
Myeong Hee KIM ; Choong Hwan CHA ; Dongheui AN ; Sung Eun CHOI ; Heung Bum OH
The Korean Journal of Laboratory Medicine 2008;28(2):144-150
BACKGROUND: Hepatitis B virus (HBV) DNA quantification is necessary for starting and monitoring of antiviral therapy in patients with chronic hepatitis B. This study was intended to assess the clinical performance of Abbott RealTime HBV Quantification kit (Abbott Laboratories, USA). METHODS: The performance was evaluated in terms of precision, linearity, detection sensitivity, cross-reactivity, and carry-over. A correlation with the Real-Q HBV Quantification kit (BioSewoom Inc., Korea) was also examined using serum samples from 64 patients diagnosed with chronic hepatitis B and underwent lamivudine therapy in Asan Medical Center. We verified the trueness of the system by comparing the outputs with the assigned values of the BBI panel (BBI Diagnostics, USA). RESULTS: Within-run and between-run coefficients of variation (CV) were 3.56-4.71% and 3.03-4.98%, respectively. Linearity was manifested ranging from 53 to 10(9) copies/mL and the detection sensitivity was verified to be 51 copies/mL. None of hepatitis C virus showed cross-reactivity. No cross-contamination occurred when negative and positive samples were alternatively placed in a row. It showed a good correlation with the Real-Q HBV (r2=0.9609) and the test results for the BBI panel were also well agreed to the assigned values (r2=0.9933). CONCLUSIONS: The performance of Abbott RealTime HBV Quantification kit was excellent; thus, it should be widely used in starting and monitoring of antiviral therapy in Korean patients with chronic hepatitis B.
Computer Systems
;
DNA, Viral/*blood
;
Hepatitis B virus/genetics/*isolation & purification
;
Hepatitis B, Chronic/*virology
;
Humans
;
Polymerase Chain Reaction/*methods
;
Reagent Kits, Diagnostic
;
Viral Load/*methods
9.Exploration of an Efficient Simultaneous Molecular Detection Method of HIV, HCV, and Syphilis from a Single Dried Blood Spot.
Jie Qiong MA ; Qing Qing XU ; Lin HE ; Xiao Xia HE ; Kai CHEN ; Yue Hua WANG ; Wen Ge XING ; Yan JIANG
Biomedical and Environmental Sciences 2021;34(4):257-264
Objective:
The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one dried blood spot (DBS) as an alternative sample to plasma.
Method:
A total of 571 paired DBS/plasma samples were collected from men who have sex with men (MSM) and injection drug users (IDUs), and serological and molecular assays were performed. Using plasma results as the reference standard, the performance of DBS tests for HIV-1 RNA, HIV-1 DNA, and HCV RNA was evaluated. Pearson's correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.
Results:
Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA, five samples (5/32) were not detectable in DBS, while measurable HIV-1 RNA levels were present in plasma (1.44 to 3.99 log
Conclusion
The performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one DBS was acceptable. DBS, as an alternative sample to plasma, may be a viable option for the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.
DNA, Viral/analysis*
;
Diagnostic Tests, Routine/methods*
;
Dried Blood Spot Testing/methods*
;
HIV Infections/diagnosis*
;
HIV-1/isolation & purification*
;
Hepacivirus/isolation & purification*
;
Hepatitis C/diagnosis*
;
RNA, Viral/analysis*
;
Sensitivity and Specificity
;
Specimen Handling/methods*
;
Syphilis/diagnosis*
;
Treponema pallidum/isolation & purification*
10.Isolation, identification and sequence analyses of dengue virus type 2 strain GD19/2001.
Rui-wen REN ; Mei-yu FANG ; Wen-yan HONG ; Bao-ming HUANG ; Lian-hua JIANG ; Jian-wei LIU ; Xiao-dong TIAN ; Gang-feng CHENG
Chinese Journal of Epidemiology 2003;24(4):288-290
OBJECTIVETo identify the virus isolated from Jiangmen, Guangdong province and to discuss the possible origin.
METHODSUsing characteristics of indirect fluorescent antibody tests (IFA), reverse transcription-polymerase chain reaction (RT-PCR), mouse neurovirulence and cell culture to identify the isolated virus. According to the nature of dengue virus type 2 NGC strain, two pairs of primers were designed. The structural protein gene of isolated dengue virus type 2 strain was then amplified by RT-PCR, cloned into pMD18-T vector and sequenced.
RESULTSTwenty-two of 37 serum samples showed a positive reaction to dengue antibody IgG, and 36 of 37 with IgM with the highest antibody titer 1:640. Ten samples were resulted in a cytopathy on C6/36 cells and showed a neurovirulence in suckling mice when inoculated intracerebrally. The structural gene of new isolate GD19/2001 containing 2 325 nucleotides which encoded 774 amino acids. Data on nucleotide homology were 98%, 96%, 94%, 94%, 92%, 92%, 92% and 91% compared with TSV01, GD06/93, NGC and 44, ThNH81/93, 04 and GD08/98, and S1 respectively.
CONCLUSIONThe isolated virus from Jiangmen, Guangdong province belonged to dengue virus type 2, which might come from Australia.
Animals ; Antibodies, Viral ; blood ; China ; epidemiology ; DNA, Viral ; genetics ; Dengue ; epidemiology ; virology ; Dengue Virus ; genetics ; immunology ; isolation & purification ; Fluorescent Antibody Technique ; Humans ; Polymerase Chain Reaction ; RNA, Viral ; genetics ; Sequence Analysis, DNA