Kết Quả Giải Trình Tự Acid Amin Protein E Chủng Giống Gốc Bv-Msv-0210 Và Chủng

660 670 680 690 700

BV-MSV-021 GTTTCATGAC CTCGCTCTCC CTTGGACGCC CCCTTCGAGC ACAGCGTGGA

BV-WSV-031 GTTTCATGAC CTCGCTCTCC CTTGGACGCC CCCTTCGAGC ACAGCGTGGA

Beijing-Ka GTTTCATGAC CTCGCTCTCC CTTGGACGCC CCCTTCGAGC ACAGCGTGGA

....|....| ....|....| ....|....|....|....| ....|....|

710 720 730 740 750

BV-MSV-021 GAAACAGAGA ACTCCTCATG GAATTTGAAG AGGCGCACGC CACAAAACAG BV-WSV-031 GAAACAGAGA ACTCCTCATG GAATTTGAAG AGGCGCACGCCACAAAACAG Beijing-Ka GAAACAGAGA ACTCCTCATG GAATTTGAAG AGGCGCACGC CACAAAACAG

....|....|....|....|....|....|....|....| ....|....|

760 770 780 790 800

BV-MSV-021 TCCGTTGTTG CTCTTGGGTC ACAGGAAGGA GGCCTCCATC AGGCGTTGGC

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BV-WSV-031 TCCGTTGTTG CTCTTGGGTC ACAGGAAGGA GGCCTCCATC AGGCGTTGGC

Beijing-Ka TCCGTTGTTG CTCTTGGGTC ACAGGAAGGA GGCCTCCATC AGGCGTTGGC

....|....| ....|....| ....|....|....|....| ....|....|

810 820 830 740 850

BV-MSV-021 AGGAGCCATC GTGGTGGAGT ACTCAAGCTC AGTGAAGTTA ACATCAGGCC

BV-WSV-031 AGGAGCCATC GTGGTGGAGT ACTCAAGCTC AGTGAAGTTA ACATCAGGCC

Beijing-Ka AGGAGCCATC GTGGTGGAGT ACTCAAGCTC AGTGAAGTTA ACATCAGGCC

....|....| ....|....|....|....| ....|....| ....|....|

860 870 880 890 900

BV-MSV-021 ACCTAAAATG CAGGCTGAAA ATGGACAAAC TGGCTCTGAA AGGCACAACC

BV-WSV-031 ACCTAAAATG CAGGCTGAAA ATGGACAAAC TGGCTCTGAA AGGCACAACC

Beijing-Ka ACCTAAAATG CAGGCTGAAA ATGGACAAAC TGGCTCTGAA AGGCACAACC

....|....| ....|....| ....|....|....|....| ....|....|

910 920 930 940 950

BV-MSV-021 TATGGTATGT GCACAGAAAA ATTCTCGTTC GCGAAAAATC CGGCGGACAC

BV-WSV-031 TATGGTATGT GCACAGAAAA ATTCTCGTTC GCGAAAAATC CGGCGGACAC

Beijing-Ka TATGGTATGT GCACAGAAAA ATTCTCGTTC GCGAAAAATC CGGCGGACAC

BV-MSV-021 TGGTCACGGA ACAGTTGTCA TTGAACTTTC ATACTCTGGG AGTGATGGCC

BV-WSV-031 TGGTCACGGA ACAGTTGTCA TTGAACTTTC ATACTCTGGG AGTGATGGCC

Beijing-Ka TGGTCACGGA ACAGTTGTCA TTGAACTTTC ATACTCTGGG AGTGATGGCC

....|....| ....|....|....|....|....|....|....|....|

1010 1020 1030 1040 1050

BV-MSV-021 CCTGCAAGAT TCCGATTGTC TCCGTTGCTA GCCTCAATGA CATGACCCCC

BV-WSV-031 CCTGCAAGAT TCCGATTGTC TCCGTTGCTA GCCTCAATGA CATGACCCCC

Beijing-Ka CCTGCAAGAT TCCGATTGTC TCCGTTGCTA GCCTCAATGA CATGACCCCC

....|....| ....|....|....|....| ....|....|....|....|

1060 1070 1080 1090 1100

BV-MSV-021 GTCGGGCGGC TGGTGACAGT GAACCCCTTC GTCGCGACTT CCAGCGCCAA

BV-WSV-031 GTCGGGCGGC TGGTGACAGT GAACCCCTTC GTCGCGACTT CCAGCGCCAA

Beijing-Ka GTCGGGCGGC TGGTGACAGT GAACCCCTTC GTCGCGACTT CCAGCGCCAA

....|....| ....|....|....|....|....|....|....|....|

1110 1120 1130 1140 1150

BV-MSV-021 CTCAAAGGTG CTGGTCGAGA TGGAACCCCC CTTCGGAGAC TCCTACATCG

BV-WSV-031 CTCAAAGGTG CTGGTCGAGA TGGAACCCCC CTTCGGAGAC TCCTACATCG

Beijing-Ka CTCAAAGGTG CTGGTCGAGA TGGAACCCCC CTTCGGAGAC TCCTACATCG

....|....| ....|....|....|....| ....|....|....|....|

1160 1170 1180 1190 1200

BV-MSV-021 TAGTTGGAAG GGGAGACAAG CAGATTAACC ACCATTGGTA CAAGGCTGGA BV-WSV-031 TAGTTGGAAG GGGAGACAAG CAGATTAACC ACCATTGGTA CAAGGCTGGA Beijing-Ka TAGTTGGAAG GGGAGACAAG CAGATTAACC ACCATTGGTA CAAGGCTGGA

....|....|....|....| ....|....|....|....| ....|....|

1210 1220 1230 1240 1250

BV-MSV-021 AGCACGCTGG GCAAAGCCTT TTCAACGACT TTGAAGGGAG CTCAAAGACT

BV-WSV-031 AGCACGCTGG GCAAAGCCTT TTCAACGACT TTGAAGGGAG CTCAAAGACT

Beijing-Ka AGCACGCTGG GCAAAGCCTT TTCAACGACT TTGAAGGGAG CTCAAAGACT

....|....| ....|....|....|....| ....|....|....|....|

1260 1270 1280 1290 1300

BV-MSV-021 GGCAGCGTTG GGCGACACAG CCTGGGACTT TGGCTCTATT GGAGGGGTCT

BV-WSV-031 GGCAGCGTTG GGCGACACAG CCTGGGACTT TGGCTCTATT GGAGGGGTCT

Beijing-Ka GGCAGCGTTG GGCGACACAG CCTGGGACTT TGGCTCTATT GGAGGGGTCT

BV-MSV-021 TCAACTCCAT AGGGAAAGCT GTTCACCAAG TGTTTGGTGG TGCCTTCAGA

BV-WSV-031 TCAACTCCAT AGGGAAAGCT GTTCACCAAG TGTTTGGTGG TGCCTTCAGA

Beijing-Ka TCAACTCCAT AGGGAAAGCT GTTCACCAAG TGTTTGGTGG TGCCTTCAGA

....|....|....|....|....|....|....|....|....|....|

1360 1370 1380 1390 1400

BV-MSV-021 ACACTCTTTG GGGGAATGTC TTGGATCACA CAAGGGCTAA TGGGGGCCCT

BV-WSV-031 ACACTCTTTG GGGGAATGTC TTGGATCACA CAAGGGCTAA TGGGGGCCCT

Beijing-Ka ACACTCTTTG GGGGAATGTC TTGGATCACA CAAGGGCTAA TGGGGGCCCT

....|....|....|....| ....|....|....|....| ....|....|

1410 1420 1430 1440 1450

BV-MSV-021 ACTACTTTGG ATGGGCATCA ACGCACGAGA CCGATCAATT GCTTTGGCCT

BV-WSV-031 ACTACTTTGG ATGGGCATCA ACGCACGAGA CCGATCAATT GCTTTGGCCT

Beijing-Ka ACTACTTTGG ATGGGCATCA ACGCACGAGA CCGATCAATT GCTTTGGCCT

....|....|....|....|....|....|....|....|....|....|

1460 1470 1480 1490 1500

BV-MSV-021 TCTTAGCCAC AGGAGGTGTG CTCGTGTTCT TAGCTACCAA TGTGCATGCT

BV-WSV-031 TCTTAGCCAC AGGAGGTGTG CTCGTGTTCT TAGCTACCAA TGTGCATGCT

Beijing-Ka TCTTAGCCAC AGGAGGTGTG CTCGTGTTCT TAGCTACCAA TGTGCATGCT

Phụ lục 2. Kết quả giải trình tự acid amin Protein E chủng giống gốc BV-MSV-0210 và chủng giống sản xuất BV-WSV-0310

....|....| ....|....|....|....|....|....|....|....|

10 20 30 40 50

Sequence JEV Beijing-Kanonji MTKKPGGPGK NRAINMLKRG LPRVFPLVGV KRVVMSLLDG RGPVRFVLAL E (ngày 10/10/19):Bei (42)2 MTKKPGGPGK NRAINMLKRG LPRVFPLVGV KRVVMSLLDG RGPVRFVLAL A (ngày 26/11/19):BV-MSV-0210 MTKKPGGPGK NRAINMLKRG LPRVFPLVGV KRVVMSLLDG RGPVRFVLAL B (ngày 26/11/19):BV-WSV-0310 MTKKPGGPGK NRAINMLKRG LPRVFPLVGV KRVVMSLLDG RGPVRFVLAL

....|....| ....|....| ....|....| ....|....|....|....|

60 70 80 90 100

Sequence JEV Beijing-Kanonji ITFFKFTALA PTKALLGRWK AVEKSVAMKH LTSFKRELGT LIDAVNKRGR E (ngày 10/10/19):Bei (42)2 ITFFKFTALA PTKALLGRWK AVEKSVAMKHLTSFKRELGT LIDAVNKRGR A (ngày 26/11/19):BV-MSV-0210 ITFFKFTALA PTKALLGRWK AVEKSVAMKHL TSFKRELGT LIDAVNKRGR B (ngày 26/11/19):BV-WSV-0310 ITFFKFTALA PTKALLGRWK AVEKSVAMKHL TSFKRELGT LIDAVNKRGR

....|....| ....|....|....|....|....|....|....|....|

110 120 130 140 150

Sequence JEV Beijing-Kanonji KQNKRGGNEG SIMWLASLAV VIACAGAMKL SNFQGKLLM TINNTDIADVI E (ngày 10/10/19):Bei (42)2 KQNKRGGNEG SIMWLASLAV VIACAGAMKL SNFQGKLLM TINNTDIADVI A (ngày 26/11/19):BV-MSV-0210 KQNKRGGNEG SIMWLASLAV VIACAGAMKL SNFQGKLLM TINNTDIADVI B (ngày 26/11/19):BV-WSV-0310 KQNKRGGNEG SIMWLASLAV VIACAGAMKL SNFQGKLLM TINNTDIADVI

....|....| ....|....| ....|....| ....|....|....|....|

160 170 180 190 200

Sequence JEV Beijing-Kanonji VIPTSKGENRC WVRAIDVGYM CEDTITYECX KLTMGNDPED VDCWCDNQE E (ngày 10/10/19):Bei (42)2 VIPTSKGENRC WVRAIDVGYM CEDTITYECP KLTMGNDPED VDCWCDNQE A (ngày 26/11/19):BV-MSV-0210 VIPTSKGENRC WVRAIDVGYM CEDTITYECP KLTMGNDPED VDCWCDNQE B (ngày 26/11/19):BV-WSV-0310 VIPTSKGENRC WVRAIDVGYM CEDTITYECP KLTMGNDPED VDCWCDNQE

....|....| ....|....|....|....|....|....|....|....|

210 220 230 240 250

Sequence JEV Beijing-Kanonji VYVQYGRCTR TRHSKRSRRS VXVQTHGESS LVNKKEAWLD STKATRYLMK E (ngày 10/10/19):Bei (42)2 VYVQYGRCTR TRHSKRSRRS VSVQTHGESS LVNKKEAWLD STKATRYLMK A (ngày 26/11/19):BV-MSV-0210 VYVQYGRCTR TRHSKRSRRS VSVQTHGESS LVNKKEAWLD STKATRYLMK B (ngày 26/11/19):BV-WSV-0310 VYVQYGRCTR TRHSKRSRRS VSVQTHGESS LVNKKEAWLD STKATRYLMK

....|....| ....|....| ....|....| ....|....|....|....|

260 270 280 290 300

Sequence JEV Beijing-Kanonji TENWIIRNPG YAFLAAILGW MLGSNNGQRV VFTILLLLVA PAXSFNCLGM E (ngày 10/10/19):Bei (42)2 TENWIIRNPG YAFLAAILGW MLGSNNGQRV VFTILLLLVA PAYSFNCLGM A (ngày 26/11/19):BV-MSV-0210 TENWIIRNPG YAFLAAILGW MLGSNNGQRV VFTILLLLVA PAYSFNCLGM B (ngày 26/11/19):BV-WSV-0310 TENWIIRNPG YAFLAAILGW MLGSNNGQRV VFTILLLLVA PAYSFNCLGM

....|....|....|....|....|....|....|....|....|....|

310 320 330 340 350

Sequence JEV Beijing-Kanonji GNRDFIEGAS GATWVDLVLE GDSCLTIMAN DKPTLDVRMI NIEASQLAEV E (ngày 10/10/19):Bei (42)2 GNRDFIEGAS GATWVDLVLE GDSCLTIMAN DKPTLDVRMI NIEASQLAEV A (ngày 26/11/19):BV-MSV-0210 GNRDFIEGAS GATWVDLVLE GDSCLTIMAN DKPTLDVRMI NIEASQLAEV B (ngày 26/11/19):BV-WSV-0310 GNRDFIEGAS GATWVDLVLE GDSCLTIMAN DKPTLDVRMI NIEASQLAEV

....|....| ....|....| ....|....| ....|....|....|....|

360 370 380 390 400

Sequence JEV Beijing-Kanonji RSYCYHASVT DISTVARCPT TGEAHNEKRA DSSYVCKQGF TDRGWGNGCG E (ngày 10/10/19):Bei (42)2 RSYCYHASVT DISTVARCPT TGEAHNEKRA DSSYVCKQGF TDRGWGNGCG A (ngày 26/11/19):BV-MSV-0210 RSYCYHASVT DISTVARCPT TGEAHNEKRA DSSYVCKQGF TDRGWGNGCG B (ngày 26/11/19):BV-WSV-0310 RSYCYHASVT DISTVARCPT TGEAHNEKRA DSSYVCKQGF TDRGWGNGCG

....|....|....|....|....|....|....|....|....|....|

410 420 430 440 450

Sequence JEV Beijing-Kanonji LFGKGSIDTC AKFSCTRKAI GRTIQPENIK YEVGIFVHGT TTSENHGNYS E (ngày 10/10/19):Bei (42)2 LFGKGSIDTC AKFSCTRKAI GRTIQPENIK YEVGIFVHGT TTSENHGNYS A (ngày 26/11/19):BV-MSV-0210 LFGKGSIDTC AKFSCTRKAI GRTIQPENIK YEVGIFVHGT TTSENHGNYS B (ngày 26/11/19):BV-WSV-0310 LFGKGSIDTC AKFSCTRKAI GRTIQPENIK YEVGIFVHGT TTSENHGNYS

....|....| ....|....| ....|....| ....|....|....|....|

460 470 480 490 500

Sequence JEV Beijing-Kanonji AQVGASQAAK FTVTPNAPSI TLKLGDYGEV TLDCEPRSGL NTEAFYVMTV E (ngày 10/10/19):Bei (42)2 AQVGASQAAK FTVTPNAPSI TLKLGDYGEV TLDCEPRSGL NTEAFYVMTV A (ngày 26/11/19):BV-MSV-0210 AQVGASQAAK FTVTPNAPSI TLKLGDYGEV TLDCEPRSGL NTEAFYVMTV B (ngày 26/11/19):BV-WSV-0310 AQVGASQAAK FTVTPNAPSI TLKLGDYGEV TLDCEPRSGL NTEAFYVMTV

....|....|....|....|....|....|....|....|....|....|

510 520 530 540 550

Sequence JEV Beijing-Kanonji GSXSFLVHRE WFHDLALPWT PPSSTAWRNR ELLMEFEEAH ATKQSVVALG E (ngày 10/10/19):Bei (42)2 GSKSFLVHRE WFHDLALPWT PPSSTAWRNR ELLMEFEEAH ATKQSVVALG A (ngày 26/11/19):BV-MSV-0210 GSKSFLVHRE WFHDLALPWT PPSSTAWRNR ELLMEFEEAH ATKQSVVALG

B (ngày 26/11/19):BV-WSV-0310 GSKSFLVHREWFHDLALPWTPPSSTAWRNRELLMEFEEAHATKQSVVALG

....|....| ....|....| ....|....| ....|....|....|....|

560 570 580 590 600

Sequence JEV Beijing-Kanonji SQEGGLHQAL AGAIVVEYSS SVKLTSGHLK CRLKMDKLAL KGTTYGMCTE E (ngày 10/10/19):Bei (42)2 SQEGGLHQAL AGAIVVEYSS SVKLTSGHLK CRLKMDKLAL KGTTYGMCTE A (ngày 26/11/19):BV-MSV-0210 SQEGGLHQAL AGAIVVEYSS SVKLTSGHLK CRLKMDKLAL KGTTYGMCTE B (ngày 26/11/19):BV-WSV-0310 SQEGGLHQAL AGAIVVEYSS SVKLTSGHLK CRLKMDKLAL KGTTYGMCTE

....|....|....|....|....|....|....|....|....|....|

610 620 630 640 650

Sequence JEV Beijing-Kanonji KFSFAKNPAD TGHGTVVIEL SYSGSDGPCK IPIVSVASLN DMTPVGRLVX E (ngày 10/10/19):Bei (42)2 KFSFAKNPAD TGHGTVVIEL SYSGSDGPCK IPIVSVASLN DMTPVGRLVT A (ngày 26/11/19):BV-MSV-0210 KFSFAKNPAD TGHGTVVIEL SYSGSDGPCK IPIVSVASLN DMTPVGRLVT B (ngày 26/11/19):BV-WSV-0310 KFSFAKNPAD TGHGTVVIEL SYSGSDGPCK IPIVSVASLN DMTPVGRLVT

....|....| ....|....| ....|....| ....|....|....|....|

660 670 680 690 700

Sequence JEV Beijing-Kanonji VNPFVATSSA NSKVLVEMEP PFGDSYIVVG RGDKQINHHW XKAGSTLGKA E (ngày 10/10/19):Bei (42)2 VNPFVATSSA NSKVLVEMEP PFGDSYIVVG RGDKQINHHW YKAGSTLGKA A (ngày 26/11/19):BV-MSV-0210 VNPFVATSSA NSKVLVEMEP PFGDSYIVVG RGDKQINHHW YKAGSTLGKA B (ngày 26/11/19):BV-WSV-0310 VNPFVATSSA NSKVLVEMEP PFGDSYIVVG RGDKQINHHW YKAGSTLGKA

....|....|....|....|....|....|....|....|....|....|

710 720 730 740 750

Sequence JEV Beijing-Kanonji FSTTLKGAQR LAAXGDTAWD FGSIGGXFNS IGKAVHQVFG GAFRTLFGGM E (ngày 10/10/19):Bei (42)2 FSTTLKGAQR LAALGDTAWD FGSIGGVFNS IGKAVHQVFG GAFRTLFGGM A (ngày 26/11/19):BV-MSV-0210 FSTTLKGAQR LAALGDTAWD FGSIGGVFNS IGKAVHQVFG GAFRTLFGGM B (ngày 26/11/19):BV-WSV-0310 FSTTLKGAQR LAALGDTAWD FGSIGGVFNS IGKAVHQVFG GAFRTLFGGM

....|....| ....|....| ....|....| ....|....|...

760 770 780 790

Sequence JEV Beijing-Kanonji SWITQGLMGA LLLWMGINAR DRSIALAXLA TGGVLVFLAT NVHA E (ngày 10/10/19):Bei (42)2 SWITQGLMGA LLLWMGINAR DRSIALAFLA TGGVLVFLAT NVHA A (ngày 26/11/19):BV-MSV-0210 SWITQGLMGA LLLWMGINAR DRSIALAFLA TGGVLVFLAT NVHA B (ngày 26/11/19):BV-WSV-0310 SWITQGLMGA LLLWMGINAR DRSIALAFLA TGGVLVFLAT NVHA

ASSESSMENT OF GENE AND PROTEIN SIMILARITIES OF BEIJING-1 VACCINE PRODUCING STRAIN WITH JAPANESE ENCEPHALITIS VIRUS

STRAINS CIRCULATING IN VIETNAM

Nguyen Thi Ly1, Huynh Thi Phuong Lien2, Do Tuan Dat2, Nguyen Kim Bach1, Nguyen Hoang

Tung1, Ha Thi Thu3, Dinh Duy Khang3,

1National Institute for Control of Vaccines and Biologicals (NICVB) 2Vaccine and Biological Products One Member Limited Company (VABIOTECH) 3Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST)

To whom correspondence should be addressed. E-mail: khangvspt@ibt.ac.vn Received: 13.8.2020

Accepted: 15.12.2020 SUMMARY

Since 2006, the inactivated Japanese encephalitis vaccine has been studied and produced by the VABIOTECH company from the Beịing-1 strain on Vero cells. Vaccines produced from the materseed and working seed virus have been evaluated at laboratory and clinical scale in humans. The results showed that the vaccine was safe and created 100% protective antibodies after the booster dose. To officially put this vaccine into production and mass use, the master seed virus BV-MSV0210 and working seed virus BV-WSV-0310 with the reference standard strain JEV Beijing-Kanonji has been tested for genetic stability. By the method of Sanger sequencing and genetic analysis software, we have evaluated the similarity of nucleotide and proteinsequencesof the E antigenencoding gene. The results showed that the seed virus similarity of amino acids and nucleotides is 100% compared with the reference strain. Thus, it can be concluded, the seed virus has antigen stability. Nucleotide and amino acid gene sequences of E genomic regions of the two seed lots were compared with virus strains isolated from human, pig and mosquito in Vietnam. The results showed that the nucleotide similarity of seed virus compared with the JEV strains isolated from humans ranged from 86.67 to 97.54%; from pigs is 87.47 to 88.33%, and from mosquitoes is 86.05 to 99%. Meanwhile, the amino acid similarity of the seed virus with the JEV strains isolated from humans ranged from 96.73 to 99.02%; from pigs is 98.00 to98.40% and from mosquitoes 94.55 to 98.40%. The sequence of amino acids in the epitope producing neutralizing antibodies of the seed virus did not differ from that of the JEV strain circulating in humans isolated in 2014.

Keywords: Beijing-1 strain, Master Seed Virus strain, Working Seed Virus strain, Envelope protein gene, nucleotide homology, amino acid similarity, neutralizing antibody-produce epitopes.

INTRODUCTION

Japanese Encephalitis virus has many strains (about 290 strains have been isolated in Asia), belonging to 5 genotypes. However, not all strains can be used to produce human vaccines. In order to be selected for the production of vaccines for human use, the virus strain needs to meet many criteria as prescribed by the World Health Organization.

Currently, most strains used to produce vaccines belong to genotype 3 such as: Nakayama strain; Beijing -1; Beijing-3; SA-14-14-2 (Sharma et al., 2014; Huynh Phuong Lien et al., 2011).

According to statistics, Japan is the first research country to produce the first inactivated JE vaccine with strains of Nakayama and/or Beijing-1. However, the actual clinical research results show

that the Beijing-1 strain has superior immunity compared to the Nakayama strain. Therefore, since 1989, the production of the JE vaccine was officially switched to use the strain Beijing-1. After Japan, China also researched and produced vaccines using strains Beijing-3 and SA-14-14-2. Subsequently, Korea started using the Nakayama strain; India and Austria used the strain SA-14-14-2.

In Vietnam, in 1989 Vabiotech company researchedand used the Nakayama strain to produce inactivated JE vaccine on mouse brains. The vaccine has been put into mass use since the early 1990s up to now with very good protection results. From 2006 up to now, Vabiotech company has researched and developed a technological process to produce inactivated JE vaccines on Vero cells from Beijing-1 strain to gradually replace vaccines produced in the brain of mice today. This new vaccine has gone through a number of stages such as the establishment of a Master Seed Bank with code BV-MSV-0210 and a Working Seed Bank with code BV-WSV-0310.The assessment and monitoring of the quality of the strains produced by the time of preservation and over the quality of 10 batches of finished vaccines at the laboratory scale and on the clinical scale in humans have been carried out from 2013-2018. The results showed that the vaccine was safe and created 100% protective antibodies after the booster injection. To officially put this vaccine into production and mass use, the evaluation of the E gene region stability

(specific protective antibody-forming region) of these strains compared with the original strain and the similarity of nucleotides and amino acids with the JEV strains circulating in Vietnam is essential to confirm whether the vaccine using this batch of strains is really effective in preventing the disease from the JEV strains circulating in Vietnam. For that reason, we performed E gene sequencing and analysis of nucleotide and amino acid similarity of master seed virus (MSV) batch BV-MSV-0210 and working seed virus (WSV) batch BV-WSV-0310 with the original standard strain (Reference JEV Beijing-Kanonji) provided by Kanonji, Japan to assess genetic stability and compare with existing JEV strains in Vietnam to evaluate antigen similarity.

In this study, we present some results on the

E gene region similarity of 2 batches of Beijing1 strains being used in JE vaccine production with standard virus strains and JEV strains circulating in Viet Nam.

MATERIALS AND METHODS

The object of study is the seed bank of batch BV-MSV-0210 and the seed bank for production batch BV-WSV-0310. Reference JEV BeijingKanonji provided by Kanonji, Japan, is the strain used to produce 2 batches of seed strains used in research. Total RNA was extracted and purified from each sample using the QIAamp Viral RNA Mini Kit (Qiagen). E gene region was amplified by RT-PCR using OneStep PCR Kit (Qiagen) with primer pair: JEV-E-p1: