1.Application of TMA Technology in Donors' HBV-DNA Detection.
Ben-Chun REN ; Xiao-Zhen ZHOU ; Xiao-Fu ZHUO
Journal of Experimental Hematology 2019;27(6):1998-2002
OBJECTIVE:
To understand the infection of hepatitis B virus(HBV) in blood donors, and to evaluate the effectiveness and necessity of TMA technology for HBV-DNA screening in blood donors.
METHODS:
Using the ELISA/NAT model, routine serology test and NAT were performed in the 169 160 donors,including voluntary blood donors and some of donors returned to donor team. For some donors with test positive NAT, nucleic acid identification test was performed. And the HBsAg neutralized and confirmed assay would conduct in blood donors with unilateral HBsAg positive and HBV-DNA negative result.
RESULTS:
Among 169 160 donation cases-times, the donors of bilateral positive of HBsAg detection was 803, accounted for 0.476%; donors of unilateral positive was 243, accounted for 0.144%. For 40 specimens with HBV-DNA negative, unilateral HBsAg positive, the neutralization and confirmed assay was performed.In result, only 4 specimens were confirmed to be HBsAg positive, the confirmed positive rate was 10%. Among detected 1003 specimens with HBV-DNA positive specimens, both HBsAg and HBV-DNA positive were 739, the consistency rate between 2 kinds of detection was 73.7%. The comparision of positive rate detected by using 3 kinds of reagents showed that there were statistical differences (P<0.05); moreover, there were statistical difference in positive rate detected by using Murex reagent and In Tec reagent (P<0.0125). The comparison of detected rate of HBsAg and HBV-DNA during March 2016-February 2017 showed no statistical difference (P>0.05). Among 60 blood donors with HBsAg and HBV-DNA who has retured to the donor team, 1 donor presented the transformation of HBsAg from negative to positive, suggesting the HBV infection of window period, HBsAg of the other 59 was negative. The detection of HBV-DNA showed that the HBV-DNA in 28 donors was negative, and the HBV-DNA in 31 donors was positive, 1 donor showed HBV-DNA was uncertain.
CONCLUSION
The routine TMA technology combined with ELISA HBsAg can effectively shorten the window period for detection of HBV infection, effectively detect the occult HBV infection, and reduce the potential risk of hepatitis B spread due to blood transfusion.
Blood Donors
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DNA, Viral
;
Hepatitis B
;
Hepatitis B Surface Antigens
;
Hepatitis B virus
;
genetics
;
Humans
2.Study on Reentry Evaluation Mode for Blood Donors Used to be HBV Reactive in Jiangsu Province.
Wen-Jia HU ; Ni-Zhen JIANG ; Shao-Wen ZHU ; Hong LIN
Journal of Experimental Hematology 2022;30(1):264-269
OBJECTIVE:
To evaluate the risk of reentry in HBV reactive blood donors and feasibility of HBV reentry strategy.
METHODS:
HBsAg+ or HBV DNA+ donors who had been quarantined for more than 6 months in Jiangsu Province could propose for reentry application. Blood samples were routinely screened by dual-ELISA for HBsAg, anti-HCV, HIV Ab/Ag, and anti- Treponema pallidum and those non-reactive ones were tested by minipool nucleic acid testing (NAT) for three times. To identify occult HBV donors, samples of NAT non-reactive were further tested by electrochemiluminescence immunoassay (ECLIA) for HBV seromarkers (including HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb). Donors of only 4 ECLIA patterns were accepted to reentry, including all 5 HBV seromarkers negative, anti-HBs only but having history of hepatitis B vaccine injection, HBcAb only, HBsAb+ / HBcAb+ with HBsAb more than 200 IU/L. Additionally, the detection rate of HBV infection was compared between routine screening mode and ECLIA, as well as the reentry qualified rate of HBsAg+ and HBV DNA+ blood donors.
RESULTS:
From Oct. 2016 to Aug. 2019, a total of 737 HBV reactive donors had applied for reentry, including 667 HBsAg+ reactive and 70 HBV DNA+ reactive donors. Among 3 screening methods, the highest HBV detection rate (43.15%, 318/737) was observed on ECLIA, while only 4.75% (35/737) on ELISA and 3.12% (23/737) on NAT, respectively. Among 4 qualified patterns of HBV serological markers, the highest proportion was found in the all negative group (22.90%, 155/677), followed by the group with HBsAb+ only and history of hepatitis B vaccine injection (19.35%, 131/677), and the median concentration of HBsAb was 237.7 IU/L. The unqualified rate of HBV DNA+ donors was 82.86%, which was significantly higher than 47.98% of HBsAg+ donors.
CONCLUSION
Routine screening tests merely based on ELISA and NAT could miss occult HBV donors and may not be sufficient for blood safety. HBsAb concentration and vaccine injection history should be included in the evaluation of HBV reactive donors who intend to apply for reentry. There is a relatively larger residual risk of occult HBV infection in blood donors quarantined for HBV DNA reactive.
Blood Donors
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DNA, Viral
;
Hepatitis B
;
Hepatitis B Surface Antigens
;
Hepatitis B virus/genetics*
;
Humans
3.Preliminary study on genotype of hepatitis B virus detected from Tibetans in China.
Yan-qing XU ; Yong-dong ZHOU ; Sheng-li BI
Chinese Journal of Experimental and Clinical Virology 2005;19(2):118-120
<b>OBJECTIVEb>To determine the main genotype of hepatitis B virus (HBV) detected from Tibetans in China and provide basic data for hepatitis control and prevention.
<b>METHODSb>The S gene and C gene were amplified by PCR from the sera of HBsAg positive Tibetans. After sequencing, the gene sequences were analyzed and the phylogenetic trees were drawn by the software MEGA3.
<b>RESULTSb>In trees based on S gene, the sequences of most samples clustered at genotype D, while in trees based on C gene, the sequences of all samples clustered at genotype C.
<b>CONCLUSIONb>The dominant genotype of HBV detected from Tibetans in China is a C/D hybrid.
Genotype ; Hepatitis B ; blood ; epidemiology ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B Surface Antigens ; blood ; genetics ; Hepatitis B virus ; classification ; genetics ; immunology ; Humans ; Phylogeny ; Tibet ; epidemiology
4.Comparision of the serotypes and genotypes of hepatitis B virus among HBsAg positive hepatitis B patients in a northern city and a southern city of China.
Zhi Wu LÜ ; Jun XU ; Ji-Mei YIN ; Wei XU
Chinese Journal of Hepatology 2005;13(2):95-100
China
;
Female
;
Genotype
;
Hepatitis B
;
blood
;
virology
;
Hepatitis B Surface Antigens
;
blood
;
Hepatitis B virus
;
classification
;
genetics
;
Humans
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Male
;
Serotyping
6.Occult hepatitis B virus infection.
Chang-yun CHEN ; Jian-rui WU ; Xin-yu ZHU ; Shuo-yi WANG
Chinese Journal of Hepatology 2005;13(11):873-875
7.Comparison of hepatitis B virus serotype and genotype among HBsAg positive hepatitis B patients in a northern and a southern city of China.
Jun XU ; Qi-xin WANG ; Chun-lei FAN ; Dong JIANG ; Ruo-bing LI ; Xu CONG ; Ran FEI ; Hong-song CHEN ; Lai WEI ; Yu WANG
Chinese Journal of Experimental and Clinical Virology 2003;17(4):327-329
<b>OBJECTIVEb>To understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China.
<b>METHODSb>Using polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China.
<b>RESULTSb>Comparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang.
<b>CONCLUSIONb>The results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar.
China ; DNA, Viral ; genetics ; Genotype ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; Serotyping
8.Quantitative analysis of HBV DNA amplified products with microtiter hybridization.
Quan ZHANG ; Jing-juan DING ; Xiao-hui MIAO
Chinese Journal of Experimental and Clinical Virology 2003;17(1):39-41
<b>BACKGROUNDb>To establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).
<b>METHODSb>The products of PCR using primers pre-labeled with biotin were hybridized with the capture probes that were immobilized on the microtiter strips and then bond with Sav-Ap. The quantity of DNA was detected by measuring the yellow color at 405 nm wave length.
<b>RESULTSb>Totally 125 sera from patients with hepatitis B were tested for HBV DNA by this method,the sera were also tested for HBV immunological markers by solid phase radio immuno-assay (SPRIA). The HBV positive rate with PCR-ELISA was 64.9% (24/37) in samples which were positive for HBsAg, HBeAg and HBcAb; and 34.2% (13/38) in sera which were positive for HBsAg, HBeAb and HBcAb; in sera positive for HBsAg and HBcAb or only HBcAb, the positive rate was 6.7% (1/15) and 5.9% (2/34) respectively.
<b>CONCLUSIONSb>The PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.
DNA Probes ; DNA, Viral ; analysis ; biosynthesis ; genetics ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; Sensitivity and Specificity
9.Advance in research of hepatitis B virus pre-S1 antigen.
Chinese Journal of Hepatology 2004;12(12):765-766
10.Gene chip analysis of mutation of HBV-DNA open reading frame.
Shi-jun CHEN ; Wen-jun DU ; Yan-qin LU ; Wei XU ; Yong AN
Chinese Journal of Experimental and Clinical Virology 2004;18(4):373-376
<b>OBJECTIVEb>To detect the mutation of HBV-DNA open reading frame (ORF), provide evidences for clinical diagnosis and treatment.
<b>METHODSb>By using gene chip technique, HBV-DNA was amplified by PCR, incorporated with fluorescence, hybridized with oligonucleotide to detect the gene sequence of DNA by computer analysis and observe the natural mutation of HBV-DNA ORF.
<b>RESULTSb>The mutations of HBV-DNA ORF existed widespread. The rates of mutation on Pre-C 1896, PreC 1814, BCP 1762, BCP 1764, P 528, P 552MI, P 552MV were 23.5%, 3.9%, 55.9%, 53.9%,39.2%, 38.2%, 10.8%, respectively.
<b>CONCLUSIONb>The gene chip technique possesses extremely high sensitivity and reliability,it is one of the effective methods to detect gene mutation. The mutation of HBV-DNA has important influence on the stability and progress of the disease, and on the judgement of prognosis.
DNA, Viral ; genetics ; Hepatitis B Core Antigens ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; genetics ; immunology ; Humans ; Mutation ; Oligonucleotide Array Sequence Analysis ; methods ; Open Reading Frames ; genetics