1.Development of a multiplex polymerase chain reaction system for screening low-frequency blood group antigens K and Ytb.
Li XIE ; Yunlei HE ; Xijiang KURE ; Luyi YE ; Ziyan ZHU
Chinese Journal of Medical Genetics 2014;31(1):78-81
OBJECTIVEA multiplex PCR system for screening rare blood group antigens K and Yt(b) was constructed to study the distribution of the two blood groups in a Uygur population in Xinjiang, China.
METHODSSequence-specific primers (SSP) were designed based on single nucleotide polymorphism sites of KEL and ACHE alleles encoding the two blood group antigens. The system was designed for simultaneously detecting the two antigens by optimizing the PCR reaction. Three hundred and sixty-two randomly selected healthy individuals were screened. Products of PCR were further analyzed for heterozygosity.
RESULTSThe system was set up successfully. No KK sample was identified and 9 K+ k+ , 41 Yt (a+ b+ ), 4 Yt (a- b+ ) were found among the 362 samples.
CONCLUSIONThe established PCR-SSP based multiple PCR system is efficient to screen the rare blood group antigens K and Yt(b). The information of rare blood donors obtained from the screening can be used to improve the capability of compatible transfusion.
Antigens, Bacterial ; genetics ; Antigens, Surface ; genetics ; Blood Donors ; Blood Group Antigens ; genetics ; Blood Transfusion ; methods ; China ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide
2.Application of multiplex PCR for the screening of genotyping system for the rare blood groups Fy(a-), s-,k-,Di(b-) and Js(b-).
Wei JIAO ; Li XIE ; Hailan LI ; Jiao LAN ; Zhuning MO ; Ziji YANG ; Fei LIU ; Ruiping XIAO ; Yunlei HE ; Luyi YE ; Ziyan ZHU
Chinese Journal of Medical Genetics 2014;31(2):242-246
OBJECTIVETo screen rare blood groups Fy(a-), s-, k-, Di(b-) and Js(b-) in an ethnic Zhuang population.
METHODSSequence-specific primers were designed based on single nucleotide polymorphism (SNP) sites of blood group antigens Fy(b) and s. A specific multiplex PCR system I was established. Multiplex PCR system II was applied to detect alleles antigens Di(b), k, Js(b)1910 and Js(b) 2019 at the same time. The two systems was were used to screen for rare blood group antigens in 4490 randomly selected healthy donors of Guangxi Zhuang ethnic origin.
RESULTSWe successfully made the multiplex PCR system I. We detected the rare blood group antigens using the two PCR system. There are five Fy(a-), three s(-), two Di(b-) in 4490 Guangxi zhuang random samples. The multiplex PCR system I has achieved good accuracy and stability. With multiplex PCR systems I and II, 4490 samples were screened. Five Fy(a-), three s(-) and two Di(b-) samples were discovered.
CONCLUSIONMultiplex PCR is an effective methods, which can be used for high throughput screening of rare blood groups. The rare blood types of Guangxi Zhuang ethnic origin obtained through the screening can provide valuable information for compatible blood transfusion. Through screening we obtained precious rare blood type materials which can be used to improve the capability of compatible infusion and reduce the transfusion reactions.
Blood Group Antigens ; genetics ; Duffy Blood-Group System ; genetics ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Receptors, Cell Surface ; genetics
3.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
4.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
5.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
OBJECTIVETo determine the main genotype of hepatitis B virus (HBV) detected from Tibetans in China and provide basic data for hepatitis control and prevention.
METHODSThe 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.
RESULTSIn 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.
CONCLUSIONThe 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
6.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
;
Male
;
Serotyping
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
OBJECTIVETo understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China.
METHODSUsing 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.
RESULTSComparison 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.
CONCLUSIONThe 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
9.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
10.Advance in research of hepatitis B virus pre-S1 antigen.
Chinese Journal of Hepatology 2004;12(12):765-766