OBI. ELISA HBsAg - OBI. HBsAg + B V14G /A Y161F /S V168A P217L C E2G /A /V T118R /K /A /M P127T /L /H /S E164D /G L175S S174N
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1 Chin J Blood Transfusion Jan 2015 Vol. 28 No. 1 * pre-s /S OBI pre-s /S ELISA HBsAg - HCV -HIV -TP HIV /HBV /HCV NAT PCR OBI pre-s /S HBsAg + Genbank OBI / OBI pre-s /S B 6 C 34 D 1 OBI S P B = P C = B V14G /A Y161F /S V168A P217L C E2G /A /V T118R /K /A /M P127T /L /H /S E164D /G L175S S174N OBI HBV S OBI OBI pre-s /S R R Q503 A X Analysis of pre-s /S gene in occult hepatitis B virus infection from blood donors in Dalian China WANG Dong 1 DENG Xuelian 1 ZHOU Lu 1 FANG Linlin 2 SONG Liuwei 2 YUAN Quan 2 AN Wanxin Dalian Blood Center Dalian China 2. National Institute of Diagnostics and Vaccine Development in Infectious Diseases School of Life Science Xiamen University. Corresponding author AN Wanxin. Abstract Objective To understand the prevalence of occult infection of hepatitis B virus OBI and the mutations of pre-s /S gene from blood donors in Dalian. Methods From December 2 nd 2010 to May 31 st 2013 samples from blood donors in Dalian Blood Center were screened with enzyme-linked immunosorbent assay ELISA for HBsAg anti-hcv anti- HIV and anti-tp and triplex nucleic acid detection NAT for HIV RNA HBV DNA and HCV RNA. A follow-up was conducted on donors who were tested negative by ELISA but reactive by NAT. OBI was determined by testing the specimens from the follow-up or archive with serological markers of hepatitis B virus identification examinations quantitative and semi-nested PCR. The sequences of pre-s /S gene of OBIs and the control group were analyzed HBsAg + Genbank. Results blood samples were screened 69 of which were OBIs. The prevalence rate of OBIs was / The pre-s /S gene sequences were analyzed in forty-one cases of OBI 6 were type B 34 were type C and 1 was type D. Comparing the amino acid sequences between the experimental and control groups there were statistical significance in the S region of OBI P B = P C = The main mutation sites of amino acids for type B were V14G /A Y161F /S V168A and P217L while for type C were E2G /A /V T118R /K /A /M P127T /L /H /S E164D /G L175S and S174N. Conclusion There were multiple mutation loci on the amino acid sequence of OBI from blood donors in Dalian. A particular relationship existed between these mutations in S region from HBV genome and the mechanism of OBI which was influenced by genotypes. Key words occult hepatitis B virus infection pre-s /S gene mutation in amino acid sequence hepatitis B virus blood donors Dalian occult hepatitis B virus infection OBI HBV DNA HBV DNA HBV DNA HBsAg < 200 IU /ml 1 OBI HBV doi /j. cjbt. issn x HBsAg OBI * HBV OBI anwx560129@ 163. com nucleic acids technology NAT
2 Chin J Blood Transfusion Jan 2015 Vol. 28 No OBI HBV 2 50 μl 2 μl OBI PCR pre-s /S OBI DNA Tiangen OBI pmd18-t pre-s /S Invitrogen OBI ml 3 EDTA-K NAT NCBI < 4 h g 15 min 2 http / /www. ncbi. nlm. nih. gov /projects /genotyping / 3 form page. cgi HBV Genbank ht tp / /www. ncbi. nlm. nih. gov /nuccore / term = HBV + CHI- ml JF C n = 20 AB DQ HBsAg -HCV EU EU FJ FJ FJ HIV FJ FJ FJ GQ HQ TP JF JQ JQ MEGA 5. 2 AT2 Hamilton RSP200 EVO nonparametric FAME 24 /20 Hamilton 2 Mann-Whitney test OBI 1 HBV HCV HIV P Cobas s 201 Procleix Tigris HBV NAT + ELIAS OBI Procleix Tigris Cobas Taqman HBV DNA + HBsAg - Cobas Taqman HBV DNA 1. 5 HBV HBV NAT 69 OBI 1 DNA + HBsAg - -HBc / OBI HBsAg -HBs HBeAg -HBe Cobas e ± pre-s /S NAT + ELIAS P < HBV DNA 2. 2 OBI pre-s /S 69 OBI HBV pre-s /S PCR semi-nested PCR pre-s /S 41 B 6 C 34 D Kp2356sF 5'-TCCgCTAgAgATCTgg- TACCATggCAggTCCCCTAgAAgAAgAACT-3' Hd1627R 5'- TCTTATCTAgAAgCTTgCgTTCACggTggTCTCCATgC-3' 1 20 μl DNA 5 μl 0. 4 nt DL28 pre-s2 21 nt mmol /L dntp 0. 2 μmol /L U /μl LA Taq Takara 95 4min s s 72 3 min min Kp2356sF Hd1599sR 5'-TCTTATCTAgAAgCTTACAgAggTgAAgCgAAgTgCACACgg-3' 1. 7 OBI OBI 1 NAT + ELIAS HBV DNA HBV HBV PCR NA HBV B n = 5 EU FJ FJ HQ ELISA 2 HBsAg -HCV -HIV -TP DQ DQ EU EU EU SPSS OBI ± d OBI B DL10 pre-s2 15 nt 2-16 C DL40 pre-s1 6 nt DL57 pre-s DL112 pre-s1 nt 3157 C T HBsAg + D 1 40
3 Chin J Blood Transfusion Jan 2015 Vol. 28 No. 1 B C S P < < B V14G /A 3 / 6 Y161F /S 3 /6 V168A 3 /6 P217L 3 /6 C E2G /A /V 8 /34 T118R /K /A /M 13 /34 P127T /L /H /S 8 / 34 E164D /G 8 /34 L175S 9 /34 S174N 8 / OBI % OBI % n = n = OBI u = P < OBI IU/mL PCR -HBs IU/L -HBc HBeAg COI -HBe d 1 DL001 HBV < 20 + C DL003 HBV ND - / DL004 HBV < 20 + C DL005 HBV ND - / DL006 HBV < 20 + D DL009 HBV < 20 + C DL010 HBV < 20 + B DL014 - ND + / DL022 HBV / + C DL025 HBV / - / DL027 HBV < 20 + B DL028 HBV < 20 + C DL029 HBV ND + / DL030 HBV < 20 + C DL032 HBV < 20 + B DL033 HBV C DL040 HBV < 20 + C DL041 HBV < 20 + C DL042 HBV < 20 - / DL043 HBV < 20 + C DL046 HBV ND + B DL047 HBV / - / DL048 HBV < 20 + C DL049 HBV < 20 + C DL051 HBV ND + / DL052 HBV < 20 + C DL054 HBV C DL055 HBV < 20 + C DL057 HBV < 20 + C DL060 HBV < 20 + C DL061 HBV < 20 + C DL064 - ND + C DL067 HBV < 20 + C DL068 HBV < 20 + C DL069 HBV < 20 + C DL070 HBV ND + / DL072 HBV < 20 + B DL076 HBV / + / DL078 HBV / + / DL079 - < 20 - / DL081 - ND + / DL082 HBV < 20 + C DL083 HBV ND + / DL084 HBV C DL085 HBV ND - / DL088 HBV C DL090 HBV < 20 + C DL091 - ND + C DL093 HBV ND + C DL096 - ND + / DL097 HBV ND + / DL098 - ND + /
4 Chin J Blood Transfusion Jan 2015 Vol. 28 No IU/mL PCR -HBs IU/L -HBc HBeAg COI -HBe d 53 DL099 HBV < 20 + C DL100 HBV < 20 - / DL102 HBV C DL104 - ND + / DL107 HBV < 20 + B DL108 - ND + / DL109 HBV C DL110 HBV < 20 + / DL111 HBV ND + C DL112 HBV < 20 + C DL113 HBV < 20 + C DL115 - ND + / DL116 HBV < 20 + / DL120 - ND + / DL123 - ND + / DL127 HBV < 20 + / DL128 - / + / NAT 1 ND = not detected 1 1 OBI S
5 Chin J Blood Transfusion Jan 2015 Vol. 28 No. 1 3 OBI HBsAg + pre-s /S B C OBI B n = 6 HBsAg + n = 5 Mann-Whitney U P OBI C n = 34 HBsAg + n = 20 Mann-Whitney U P Pre-S Pre-S S MHR OBI MHR S 3 OBI 1 < 20 IU /ml 59. 4% 41 / / OBI 2-5 OBI 18 -HBc % 62 /69 -HBs 19 HBV 100 IU /L 60 /69 OBI OBI OBI n = 6 n = 5 pre-s1 P < Raimondo R Allain JP Brunetto MR et al. Statements from the S B C OBI 1 S OBI S G119R C121Y C124R /Y /S I /T126N /S / L Q129R / N / P / H G130N T131N M133I / T S136P C137W C139R T140I K141E D144A /E /G G145R /A /D / K G145R G145R 1 HBsAg Microbiol S HBsAg 1 OBI 82. 6% 57 /69 OBI PCR 41 pre-s /S C B 2 OBI C HBsAg % 34 / % 6 /41 1 D C OBI 14 HBV B C OBI C D B Genbank HBV C HBV pre-s /S pre-s1 pre-s2 S 3 HBV DNA + HBsAg + 3 5' OBI S HBsAg OBI S OBI OBI S MHR AA a AA OBI 22 OBI HBV OBI MHR OBI HBV S B C OBI D A2 A B Taormina expert meeting on occult hepatitis B virus infection. J Hepatol Zheng X Ye X Zhang L et al. Characterization of occult hepatitis B virus infectio from blood donors in China. J Clin Microbiol Yuan Q Ou SH Chen CR et al. Molecular characteristics of occult hepatitis B virus from blood donors in southeast China. J Clin OBI 4 Candotti D Lin CK Belkhiri D et al. Occult hepatitis B infection in blood donors from South East Asia molecular characterisati- OBI HBV DNA + HBsAg - on and potential mechanisms of occurrence. Gut OBI G145R OBI OBI de la Fuente RA Gutiérrez ML Garcia-Samaniego J et al. Pathogenesis of occult chronic hepatitis B virus infection. World J Gas- B C S OBI troenterol
6 Chin J Blood Transfusion Jan 2015 Vol. 28 No hepatitis B virus strains derived worldwide genotypes subgeno- 8. types and HBsAg subtypes. Intervirol Allain JP Mihaljevic I Gonzalez-Fraile M et al. Infectivity of blood products from donors with occult hepatitis B virus infection. Transfusion Huang CH Yuan Q Chen PJ et al. Influence of mutations in hepatitis B virus surface protein on viral antigenicity and phenotype in occult HBV strains from blood donors. J Heptal Kim H Lee SA Kim DW et al. Naturally occurring mutations in large surface genes related to occult infection of hepatitis B virus genotype C. PLOS One e Chang MH. Hepatitis B virus infection. Semin Fetal Neonatal Med HBV HBV Norder H Courouce AM Coursaget P et al. Genetic diversity of 17 Liu Y Wang J Huang Y et al. Molecular epidemiological study of hepatitis B virus in blood donors from five Chinese blood centers. Arch Virol Milborrow HM Burneet L Lowe SB et al. Rapid method for the concentration of hepatitis B virus particles using Beckman airfuge. J Clin Pathol Sanyal D Kudesia G Corbitt G. Comparison of ultracentrifugation and polyethylene glycol precipitation for concentration of hepatitis B virus HBV DNA for molecular hybridisation tests and the relationship of HBV-DNA to HBe antigen and anti-hbe status. J Med Microbiol Allain JP Belkhiri D Vermeulen M et al. Characterization of oc- 12. S. cult hepatitis B virus strains in South African blood donors. Hepatol Candotti D Grabarczyk P Ghiazza P et al. Characterization of occult hepatitis B virus from blood donors carrying genotype A2 or 14. genotype D strains. J Hepatol pre-s /S 15. HBsAg # 2# 2# NO SNP 42 d ml / 42 d 2 10 μmol /L SNP SNP PBS 1 h ATP PS CD47 Percoll g /ml PS ± vs ± P < P > PS ± vs 2. 3 ± ± vs ± P < P > EI ATP NaCl CD ± vs ± ± vs ± ± vs ± ± vs ± ± vs ± P < P > NO SNP 3 R R doi /j. cjbt. issn x A X jxliu8122@ vip. sina. com
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