Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention
Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chickens and is distributed worldwide. Primary viral antigen is VP2. The VP2 gene contains hypervariable re-gion (VP2 HRV). Mutations in this region lead to emergence of antigenically different IBDV strains. For IBDV...
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Інститут молекулярної біології і генетики НАН України
2018
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irk-123456789-1542602019-07-07T12:38:24Z Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention Pastyria, A.S. Budzanivska, I.G. Polischuk, V.P. Molecular and Cell Biotechnologies Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chickens and is distributed worldwide. Primary viral antigen is VP2. The VP2 gene contains hypervariable re-gion (VP2 HRV). Mutations in this region lead to emergence of antigenically different IBDV strains. For IBDV prevention, vaccination is used. The efficacy of vaccination depends on the ge-netic closeness of field and vaccine strains. Aim of the study was to analyze nucleotide sequence of different vaccine and field strains of IBDV circulating in Ukrainian poultry farms. Methods. In this study 11 vaccine strains and 16 field isolates were used. RNA was extracted using a magnetic separation method, reverse transcription was carried out and PCR was performed using specific primers to the VP2 gene. Obtained amplicons were used for sequencing. Phylogenetic and amino acid analysis was performed with MEGA 6 software. Results. 11 vaccine strains formed 5 phy-logenetic clusters. Cluster I represented strains GM97, 228E and MB/20. Cluster II contained mild vaccine strains LC-75 and D78. Intermediate strains Winterfield-2512 and Lukert formed cluster III. ‘Hot’ vaccine strains MB and MB/3 formed cluster IV. Cluster V was represented by strains MB/5 and V877. After addition of 16 Ukrainian field strains the tree structure remained the same. 8 isolates clustered together with ‘hot’, 5 – with intermediate, and 3 – with mild vaccine strains. Amino acid analyses confirmed antigenic closeness among vaccine and field strains of the same cluster. Conclusion. The obtained data can be used for the vaccine selection for IBD pre-vention in each particular poultry farm of Ukraine. Вірус інфекційної бурсальної хвороби (ІБХ) викликає висококонтагіозне захворювання курчат та поширений у всьому світі. Головним антигеном є білок VP2. Ген VP2 містить гіперваріабельний регіон, мутації в якому призводять до появи нових антигенних варіантів вірусу ІБХ. Для попередження інфікування використовують вакцинацію, ефективність якої залежить від генетичної спорідненості вакцинних та польових штамів вірусу. Мета. проаналізувати нуклеотидну послідовність різних вакцинних та польових штамів вірусу ІБХ, поширених в господарствах України. Методи. У дослідженні було використано 11 вакцинних та 16 польових штамів вірусу ІБХ. РНК виділяли методом магнітної сорбції, здійснювали реакцію зворотної транскрипції та постановку ПЛР із специфічними праймерами до гену VP2. Амплікони секвенували, нуклеотидну та амінокислотну послідовність аналізували за допомогою програми MEGA 6. Результати. 11 вакцинних штамів формували 5 кластерів. Кластер І містив штами GM97, 228E та MB/20. Кластер ІІ бус сформований м’якими штамами LC-75 та D78. Середні штами Winterfield-2512 та Lukert формували кластер ІІІ. «Гарячі» штами MB та MB/3 містились у кластері IV. Кластер V містив штами MB/5 та V877. Після включення 16 польових ізолятів, виявлених в Україні, структура дерева не змінилась. Вісім з них групувалися разом з «гарячими», 5 – з середніми та 3 – з м’якими вакцинними штамами. Порівняння амінокислотних послідовностей підтвердило антигенну спорідненість штамів, що знаходились в одному кластері. Висновки. Отримані результати можуть бути використані для підбору вакцин для профілактики ІБХ в кожному окремому господарстві України. Вирус инфекционной бурсальной болезни (ИББ) вызывает высококонтагиозное заболевание цыплят и распространен во всем мире. Главным антигеном вируса является белок VP2. Ген VP2 содержит гипервариабельный регион, мутации в котором приводят к появлению новых антигенных вариантов вируса ИББ. Для предупреждения инфицирования используют вакцинацию, эффективность которой зависит от генетического родства вакцинных и полевых штаммов вируса. Цель. проанализировать нуклеотидную последовательность различных вакцинных и полевых штаммов вируса ИББ, распространенных в хозяйствах Украины. Методы. В исследовании были использованы 11 вакцинных и 16 полевых штаммов вируса ИББ. РНК выделяли методом магнитной сорбции, осуществляли реакцию обратной транскрипции и постановку ПЦР со специфическими праймерами к гену VP2. Ампликоны секвенировали, нуклеотидную и аминокислотную последовательность анализировали с помощью программы MEGA 6. Результаты. 11 вакцинных штаммов формировали 5 кластеров. Кластер I содержал штаммы GM97, 228E и MB/20. Кластер II был сформирован мягкими штаммами LC-75 и D78. Средние штаммы Winterfield-2512 и Lukert формировали кластер ІІІ. «Горячие» штаммы MB и MB/3 содержались в кластере IV. Кластер V содержал штаммы MB/5 и V877. После включения 16 полевых изолятов, выявленных в Украине, структура дерева не изменилась. Восемь из них группировались вместе с «горячими», 5 – средними и 3 – мягкими вакцинными штаммами. Сравнение аминокислотных последовательностей подтвердило антигенное родство штаммов, находящихся в одном кластере. Выводы. Полученные результаты могут быть использованы для подбора вакцин для профилактики ИБХ в каждом отдельном хозяйстве Украины. 2018 Article Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention / A.S. Pastyria, I.G. Budzanivska, V.P. Polischuk // Вiopolymers and Cell. — 2018. — Т. 34, № 1. — С. 24-31. — Бібліогр.: 15 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.00096D http://dspace.nbuv.gov.ua/handle/123456789/154260 578 en Вiopolymers and Cell Інститут молекулярної біології і генетики НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Molecular and Cell Biotechnologies Molecular and Cell Biotechnologies |
| spellingShingle |
Molecular and Cell Biotechnologies Molecular and Cell Biotechnologies Pastyria, A.S. Budzanivska, I.G. Polischuk, V.P. Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention Вiopolymers and Cell |
| description |
Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chickens and is distributed worldwide. Primary viral antigen is VP2. The VP2 gene contains hypervariable re-gion (VP2 HRV). Mutations in this region lead to emergence of antigenically different IBDV strains. For IBDV prevention, vaccination is used. The efficacy of vaccination depends on the ge-netic closeness of field and vaccine strains. Aim of the study was to analyze nucleotide sequence of different vaccine and field strains of IBDV circulating in Ukrainian poultry farms. Methods. In this study 11 vaccine strains and 16 field isolates were used. RNA was extracted using a magnetic separation method, reverse transcription was carried out and PCR was performed using specific primers to the VP2 gene. Obtained amplicons were used for sequencing. Phylogenetic and amino acid analysis was performed with MEGA 6 software. Results. 11 vaccine strains formed 5 phy-logenetic clusters. Cluster I represented strains GM97, 228E and MB/20. Cluster II contained mild vaccine strains LC-75 and D78. Intermediate strains Winterfield-2512 and Lukert formed cluster III. ‘Hot’ vaccine strains MB and MB/3 formed cluster IV. Cluster V was represented by strains MB/5 and V877. After addition of 16 Ukrainian field strains the tree structure remained the same. 8 isolates clustered together with ‘hot’, 5 – with intermediate, and 3 – with mild vaccine strains. Amino acid analyses confirmed antigenic closeness among vaccine and field strains of the same cluster. Conclusion. The obtained data can be used for the vaccine selection for IBD pre-vention in each particular poultry farm of Ukraine. |
| format |
Article |
| author |
Pastyria, A.S. Budzanivska, I.G. Polischuk, V.P. |
| author_facet |
Pastyria, A.S. Budzanivska, I.G. Polischuk, V.P. |
| author_sort |
Pastyria, A.S. |
| title |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention |
| title_short |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention |
| title_full |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention |
| title_fullStr |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention |
| title_full_unstemmed |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention |
| title_sort |
characterization of vaccine and field ibdv strains in ukraine for proper vaccine selection for disease prevention |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2018 |
| topic_facet |
Molecular and Cell Biotechnologies |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/154260 |
| citation_txt |
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention / A.S. Pastyria, I.G. Budzanivska, V.P. Polischuk // Вiopolymers and Cell. — 2018. — Т. 34, № 1. — С. 24-31. — Бібліогр.: 15 назв. — англ. |
| series |
Вiopolymers and Cell |
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2025-07-14T05:55:01Z |
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2025-07-14T05:55:01Z |
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1837600605909024768 |
| fulltext |
24
A. S. Pastyria, I. G. Budzanivska, V. P. Polischuk
© 2018 A. S. Pastyria et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Bio-
polymers and Cell. This is an Open Access article distributed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium,
provided the original work is properly cited
UDС 578
Characterization of vaccine and field IBDV strains in Ukraine for
proper vaccine selection for disease prevention
A. S. Pastyria, I. G. Budzanivska, V. P. Polischuk
ESC "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv
64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
ann.pastyria@gmail.com
Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chickens
and is distributed worldwide. VP2 is the major viral antigen. The VP2 gene contains hyper-
variable region (VP2 HRV). Mutations in this region lead to emergence of antigenically
different IBDV strains. Vaccination is used for IBDV prevention. The efficacy of vaccination
depends on similarity of field and vaccine strains. Aim. To analyze nucleotide sequences of
different vaccine and field strains of IBDV circulating in Ukrainian poultry farms. Methods.
Eleven vaccine strains and 16 field isolates were used in this study. RNA was extracted us-
ing a magnetic separation method, reverse transcription was carried out and PCR was per-
formed using specific primers to the VP2 gene. The obtained amplicons were sequenced.
Phylogenetic and amino acid analysis was performed usning the MEGA 6 software. Results.
Eleven vaccine strains formed five phylogenetic clusters. Cluster I represented strains GM97,
228E and MB/20. Cluster II contained mild vaccine strains LC-75 and D78. Intermediate
strains Winterfield-2512 and Lukert formed cluster III. ‘Hot’ vaccine strains MB and MB/3
formed cluster IV. Cluster V was represented by strains MB/5 and V877. After addition of
16 Ukrainian field strains, the tree structure remained the same. Eight isolates clustered
together with ‘hot’, five, with intermediate, and three, with mild vaccine strains. Amino acid
analyses confirmed the antigenic similarity among vaccine and field strains of the same
cluster. Conclusion. The obtained data can be used for the vaccine selection for IBD preven-
tion in poultry farms in Ukraine.
K e y w o r d s: IBDV, field strains, vaccine strains, vvIBDV, phylogenetic analysis.
Introduction
Infectious bursal disease virus (IBDV) is a
member of the Birnaviridae family Avibirna
virus genus. It is a non-enveloped virus with
two segments of double stranded RNA. The
IBDV genome is divided into segments A
(3.4 kb) and B (2.8 kb). The large segment A
encodes 4 viral proteins, the two capsid pro-
teins VP2 (48 kDa) and VP3 (32–35 kDa), the
viral protease VP4 (24 kDa), and a nonstruc-
tural protein VP5 (17–21 kDa), while the
ISSN 19936842 (online); ISSN 02337657 (print)
Biopolymers and Cell. 2018. Vol. 34. N 1. P 24–31
doi: http://dx.doi.org/10.7124/bc.00096D
mailto:ann.pastyria@gmail.com
25
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention
smaller segment B encodes VP1 (90 kDa), an
RNA-dependent RNA polymerase [1].
Virus replicates in immature IgM+ B-cells
residing in the bursa of Fabricius of young
chickens and causes an immunosuppressive
infectious bursal disease (IBD) [2]. Primary
viral antigen and structural protein is VP2.
VP2 gene contains special hypervariable re-
gion (VP2 HRV). Mutations in this region lead
to emergence of antigenically different IBDV
strains [3]. The most effective way of IBD
prevention is vaccination with the live attenu-
ated vaccines. These vaccines are divided into
three groups: “mild”, “intermediate” and in-
termediate plus or “hot” according to a degree
of their residual virulence [4]. Mild vaccines
are safe for specific pathogen free (SPF) chi-
ckens but are not very effective in the pres-
ence of high levels of maternal antibodies or
against very virulent strains of IBDV.
Intermediate and ‘hot’ vaccines are much
more effective but may induce moderate to
severe lesions in the bursa of Fabricius [5].
Choosing the appropriate vaccine is critical
for controlling IBD because of the antigenic
divergence observed among the serotype 1 vi-
ruses. Point mutations and recombination
events have contributed to the antigenic drift
and antigenic variation among IBDV strains.
In order to choose the most effective vaccine
it is necessary to analyze what kind of field
virus circulates in the farm. Vaccine strain
should be genetically and antigenically close
to the field virus [5, 6].
This study was aimed at comparing VP2
HRV of different vaccine strains and field
isolates detected in Ukraine to find the most
antigenically close vaccine strains for preven-
tion IBD in each particular farm.
Materials and Methods
In this study 11 vaccine strains (including
3 Ukrainian) and 16 Ukrainian field isolates were
used. IBDV vaccines Polimun IBD light (strain
MB/20, BioTestLab, Ukraine), Polimun IBD
(strain MB/5, BioTestLab, Ukraine), Polimun
IBD+ (strain MB/3, BioTestLab, Ukraine),
AviPro® PRECISE (strain LC-75, Loh mann
Animal Health GmbH, Germany), HIPRA-
GUMBORO® GM97 (strain GM97, Hipra, Spain)
currently used in Ukraine, are not presented in
GeneBank, that is] why they were taken from the
manufactures for the analysis of the vaccine
strains. Also, several sequences of vaccine stra-
ins were taken from GeneBank, such as:
D78 (AJ586963), 228E (AF457204), Lukert
(AY918948), V877 (AJ878882), MB (AY739669),
Winterfield-2512 (DQ355819). Ukrainian strains
were isolated from different regions as described
in previous study [7].
RNA was extracted with the use of mag-
netic separation method following supplier`s
recommendations (MagVetTM, LSI) from bur-
sa tissues samples and vaccine samples.
Reverse transcription was performed using a
commercial test kit (Reverta-L, Amplisens).
Obtained cDNA was used for nested PCR.
PCR amplification of hypervariable region of
VP2 gene was carried out with specific pri-
mers [7]. The oligonucleotide primers used in
this work designated Bur1F (5’-TCACCGTCC
TCAGCTTAC-3’ nucleotide position 587-604)
and Bur1R (5’-TCAGGATTTGGGATCAGC-
3’ nucleotide position 1212-1229) designed to
amplify the hypervariable region of VP2 gene
amplicon size – 643 bp. To increase specific-
ity and sensitivity of the reaction the second
set of primers Bur2F (5’-CGCTATAGCGCT
TGACCCAAAAA-3’, nucleotide position
26
A. S. Pastyria, I. G. Budzanivska, V. P. Polischuk
651 - 673) and Bur2R (5’-CTCACCCCAGCG
ACCGTAACGACG-3’, nucleotide position
1179-1202) designed by Kataria et al [8] were
used which allows the amplification of the in-
ner region of the first amplicon obtained after
the first round of the amplification using Bur1F
and Bur1R primers. The resulting product had
the length of 552 bp. [The] First round of am-
plification was carried out for 1 cycle at 95 °C
for 2 min, 36 cycles at 95°C for 30 s, 52 °C
for 30 s, 72 °C for 30 s, and 1 cycle at 72 °C
for 2 min. Amplicons obtained from the first
reaction were diluted by 20 times and used for
the second reaction. Thermal profile for the
second reaction was similar except the primer
annealing temperature, which was 63 °C. PCR
products were visualized in 1,5 agarosegel.
Amplicons were separated from reaction com-
ponents using the Thermo Scientific GeneJET
Gel Extraction Kit. Purified amplicons were
sequenced using forward primer (Bur2F) by
Institute of Molecular Biology and Genetics
NAS, Ukraine (3130 Genetic Analyzer,
Applied Biosystems, USA). Sequences were
analyzed using Mega 6 software [9]. Nucleotide
alignment was performed using ClustalW in-
strument. Phylogenetic analysis was performed
using neighbor-joining method [10].
Results and Discussion
426 nucleotide fragments of [the] VP2 gene of
IBDV strains were obtained and used for phy-
logenetic analysis. 11 vaccine strains formed 5
phylogenetic clusters. Cluster I represented
strains GM97, 228E and MB/20. Mean nucleo-
tide identity was 99.7%. Cluster II contained
mild vaccine strains LC-75 and D78 with nu-
cleotide identity 99.3%. Intermediate strains
Winterfield-2512 and Lukert formed cluster III
(94.5% identity). ‘Hot’ vaccine strains MB and
MB/3 formed cluster IV with 99.5% nucleotide
identity. Cluster V was represented by strains
MB/5 and V877 (98.9% identity) (Fig. 1).
To analyze phylogenetic relationship of
vaccine and field strains we added previously
described very virulent (Ukraine 1517, Ukrai-
Fig.1. Phylogenetic tree of
VP2 hypervariable region
(HRV) nucleotide sequen-
ces of 11 vaccine strains
com monly used in Ukrai-
ne. The Neighbor-Joining
consensus tree is shown.
Results of the bootstrap
test (1000 replicates) are
shown next to the branch-
es [11]. The bar represents
0.01 nucleotide substitu-
tions per site. Vaccine stra-
ins grou ped phylogeneti-
cally into mild (II), inter-
mediate (I, III, V) and
“hot” (IV) strains.
27
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention
ne 55, Ukraine 691_35_4, Ukraine 691_35_5,
Ukraine 760_45_4, Ukraine 2065, Ukraine 934,
Ukraine 964) and classical virulent (Ukrai-
ne 1853, Ukraine 38_1943, Ukraine 43_1943,
Ukraine 2045, Ukraine 58, Ukraine 691_24,
Ukraine 760_45_5, Ukraine 1147) strains iso-
lated from different regions of Ukraine to phy-
logenetic tree. It was shown that after addition
of 16 Ukrainian field strains the basic structure
of consensus tree remained the same and con-
tained the same 5 clusters (Fig. 2). 8 very
virulent field isolates clustered together with
‘hot’ and 8 classical virulent – with intermedi-
ate vaccine strains. There also were no field
strains represented in cluster III (Fig. 2).
To determine antigenic difference among
vaccine and field strains, the deduced amino
acid substitutions in the variable VP2 (vVP2)
Fig. 2. Phylogenetic tree of
hypervariable region (HRV)
nucleotide sequen ces of 11
vaccine strains and 16 field
IBDV isolates from Ukrai-
ne. The Neighbor-Joining
consensus tree is shown.
Re sults of the bootstrap test
(1000 replicates) are shown
next to the branches [11].
The bar represents 0.01 nu-
cleotide substitutions per
site. IBDV strains formed
5 clus ters. Classical virulent
IBDV strains are represent-
ed in clusters I, II and V.
Very virulent IBDV strains
are represented in cluster V.
Cluster III contains only in-
termediate vaccine strains.
28
A. S. Pastyria, I. G. Budzanivska, V. P. Polischuk
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29
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention
region (206-350 aa) [12] were analyzed (Fig. 3).
For the reference and numbering the strain
52/70 (GeneBank accession no. D00869) was
chosen [13]. The reference vvIBDV strain
UK661 (GeneBank accession no. AJ878898)
and ‘hot’ vaccine strain MB (GeneBank acces-
sion no. AY739669) were taken for comparison
with Ukrainian vvIBDV isolates. The analyzed
region included 142 amino acid residues, from
position 206 to 347. It was found that none of
the local examined isolates was of vaccine or
attenuated origin due to absence of 253-His ti-
di ne and 284-Threonine mutations that are
typically found in attenuated vaccine strains [14,
15]. Figure 3 demonstrated that 8 of the exam-
ined strains (Ukraine 1517, Ukraine 55, Ukraine
691_35_4, Ukraine 691_35_5, Ukrai ne
760_45_4, Ukraine 2065, Ukraine 934, Ukrai-
ne 964) showed the characteristic of vvIBDV
amino acid substitutions at residues 222A, 242I
(except Ukraine 760_45_4), 256I, 294I, and
299S (except Ukraine 691_35_5). 8 strains had
the serine-rich heptapeptide SWSASGS which
was found next to the second hydrophilic region
326–332 that confirmed the highly virulence
nature among the analyzed strains [14]. In five
strains [Ukraine 1517, Ukraine 55, Ukraine
691_35_4, Ukraine 691_35_5, Ukraine
760_45_4] D to N mutation in 212 aa position
was found, which was not described before. The
influence of this mutation on IBDV virulence
should be analyzed in further studies.
The comparison amino acid sequences of
very virulent isolates and vaccine strain MB
showed that in the discussed critical aa sites
MB strain is similar to the analyzed field iso-
lates. These results indicate that based on phy-
logenetic and the amino acid sequence analy-
ses vaccine strains can be chosen for preven-
tion IBD in farms. The most phylogenetically
and antigenically close to vvIBDV isolates
were MB and MB/3 strains.
Conclusion
As a result of phylogenetic analysis the vac-
cine strains studied in the research formed
5 genetic clusters. After addition of the field
strains for analysing they clustered to the most
similar vaccine strains. Deduced amino acid
analysis revealed that 8 Ukrainian isolates had
the amino acid sites common for very virulent
strains. These strains were antigenically close
to the ‘hot’ vaccine strains. Therefore, these
data can be used for the vaccine selection for
prevention IBD in each particular poultry farm.
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Характеристика вакцинних та польових
штамів вірусу ІБХ в Україні з метою
правильного підбору вакцин для профілактики
захворювання
А. С. Пастиря, І. Г. Будзанівська, В. П. Поліщук
Вірус інфекційної бурсальної хвороби (ІБХ) викликає
висококонтагіозне захворювання курчат та поширений
у всьому світі. Головним антигеном є білок VP2. Ген
VP2 містить гіперваріабельний регіон, мутації в якому
призводять до появи нових антигенних варіантів вірусу
ІБХ. Для попередження інфікування використовують
вакцинацію, ефективність якої залежить від генетичної
спорідненості вакцинних та польових штамів вірусу.
Мета. проаналізувати нуклеотидну послідовність різних
вакцинних та польових штамів вірусу ІБХ, поширених
в господарствах України. Методи. У дослідженні було
використано 11 вакцинних та 16 польових штамів ві-
русу ІБХ. РНК виділяли методом магнітної сорбції,
здійснювали реакцію зворотної транскрипції та поста-
новку ПЛР із специфічними праймерами до гену VP2.
Амплікони секвенували, нуклеотидну та амінокислотну
послідовність аналізували за допомогою програми
MEGA 6. Результати. 11 вакцинних штамів формували
5 кластерів. Кластер І містив штами GM97, 228E та
MB/20. Кластер ІІ бус сформований м’якими штамами
LC-75 та D78. Середні штами Winterfield-2512 та Lukert
формували кластер ІІІ. «Гарячі» штами MB та MB/3
містились у кластері IV. Кластер V містив штами MB/5
та V877. Після включення 16 польових ізолятів, вияв-
лених в Україні, структура дерева не змінилась. Вісім з
них групувалися разом з «гарячими», 5 – з середніми
та 3 – з м’якими вакцинними штамами. Порівняння
амінокислотних послідовностей підтвердило антигенну
спорідненість штамів, що знаходились в одному клас-
тері. Висновки. Отримані результати можуть бути ви-
користані для підбору вакцин для профілактики ІБХ в
кожному окремому господарстві України.
К л юч ов і с л ов а: вірус ІБХ, польові штами, вак-
цинні штами, високовірулентні штами, філогенетичний
аналіз.
Характеристика вакцинных и полевых
штаммов вируса ИББ в Украине с целью
правильного подбора вакцин для
профилактики болезни
А. С. Пастыря, И. Г. Будзанивская, В. П. Полищук
Вирус инфекционной бурсальной болезни (ИББ) вы-
зывает высококонтагиозное заболевание цыплят и
распространен во всем мире. Главным антигеном ви-
руса является белок VP2. Ген VP2 содержит гиперва-
риабельный регион, мутации в котором приводят к
появлению новых антигенных вариантов вируса ИББ.
Для предупреждения инфицирования используют вак-
31
Characterization of vaccine and field IBDV strains in Ukraine for proper vaccine selection for disease prevention
цинацию, эффективность которой зависит от генети-
ческого родства вакцинных и полевых штаммов виру-
са. Цель. проанализировать нуклеотидную последо-
вательность различных вакцинных и полевых штаммов
вируса ИББ, распространенных в хозяйствах Украины.
Методы. В исследовании были использованы 11 вак-
цинных и 16 полевых штаммов вируса ИББ. РНК
выделяли методом магнитной сорбции, осуществляли
реакцию обратной транскрипции и постановку ПЦР
со специфическими праймерами к гену VP2. Ампли-
ко ны секвенировали, нуклеотидную и аминокислот-
ную последовательность анализировали с помощью
программы MEGA 6. Результаты. 11 вакцинных штам-
мов формировали 5 кластеров. Кластер I содержал
штаммы GM97, 228E и MB/20. Кластер II был сфор-
мирован мягкими штаммами LC-75 и D78. Средние
штаммы Winterfield-2512 и Lukert формировали кластер
ІІІ. «Горячие» штаммы MB и MB/3 содержались в
кластере IV. Кластер V содержал штаммы MB/5 и V877.
После включения 16 полевых изолятов, выявленных
в Украине, структура дерева не изменилась. Восемь из
них группировались вместе с «горячими», 5 – средни-
ми и 3 – мягкими вакцинными штаммами. Сравнение
аминокислотных последовательностей подтвердило
антигенное родство штаммов, находящихся в одном
кластере. Выводы. Полученные результаты могут
быть использованы для подбора вакцин для профилак-
тики ИБХ в каждом отдельном хозяйстве Украины.
К л юч е в ы е с л ов а: вирус ИБХ, полевые штаммы,
вакцинные штаммы высоковирулентные штаммы,
филогенетический анализ.
Received 22.11.2017
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