Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5
Aim. The aim of our study was to follow the persistence of viral RNA in selected organs of experimentally infected with coxsackievirus (CV) B5 strains from different sources such as a patient’s sample, an environmental sample and a prototype virus strain. Methods. CD-1 mice were infected with CVB5...
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Інститут молекулярної біології і генетики НАН України
2011
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| Цитувати: | Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 / Stipalova D., Sojka M., Borsanyiova M., Badurova M., Marosova L., Sobotova Z., Bopegamage S. // Вiopolymers and Cell. — 2011. — Т. 27, № 2. — С. 162-164. — Бібліогр.: 15 назв. — англ. |
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irk-123456789-1537092019-06-15T01:25:19Z Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 Stipalova, D. Sojka, M. Borsanyiova, M. Badurova, M. Marosova, L. Sobotova, Z. Bopegamage, S. Short Communications Aim. The aim of our study was to follow the persistence of viral RNA in selected organs of experimentally infected with coxsackievirus (CV) B5 strains from different sources such as a patient’s sample, an environmental sample and a prototype virus strain. Methods. CD-1 mice were infected with CVB5 strain Faulkner the prototype, CVB5 – isolate from treated sewage waste and isolate from patient’s stool sample both identified as CVB5. The viral RNA was detected by RT-PCR using enterovirus primers specific for the non-coding 5' region. Results. We observed presence of RNA in the brain and heart of mice infected with isolate from patient’s stool at day 45 post infection (p. i.). Conclusion. We conclude that CVB5 persists in the brain and heart after oral infection of CD1 mice. The relevance of viral persistence maybe related viral origin and the genetics. Keywords: coxsackievirus B5, mouse, brain, persistence. Мета нашого дослідження полягала у тому, щоб прослідкувати, як вірусна РНК зберігається в окремих органах мишей, експериментально інфікованих деякими штамами вірусу Коксакі (ВК) B5, виділеному з різних джерел: із зразка пацієнта, навколишнього середовища і штаму прототипу вірусу. Методи. Мишей CD-1 інфікували ВКВ5 – прототипом Фолкнера і двома вірусами, ідентифікованими як ВКВ5: перший виділено з оброблених відходів стічних вод, другий – із фекалій пацієнта. Вірусну РНК виявлено методом РТ-ПЛР з використанням праймерів, специфичних для некодуючих 5'-ділянок ентеровірусів. Результаты. Ми спостерігали присутність РНК вірусу в головному мозку і серці мишей, інфікованих ізолятом із фекалій пацієнта на 45-й день після зараження (PI). Висновки. Зроблено висновок стосовно того, що ВКВ5 зберігається в мозку і серці після перорального зараження CD1 мишей. Ступінь вірусної стійкості можу бути пов’язаний з походженням вірусу та його генетикою. Ключові слова: вірус Коксакі B5, миші, мозок, стійкість. Цель нашего исследования состояла в том, чтобы проследить, как вирусная РНК сохраняется в отдельных органах мышей, экспериментально инфицированных различными штаммами вируса Коксаки (ВК) B5, выделенными из разных источников: из образца пациента, окружающей среды и штамма прототипа вируса. Методы. Мышей CD-1 инфицировали ВКВ5 – прототипом Фолкнера и двумя вирусами, идентифицированными как ВКВ5: первый выделен из обработанных отходов сточных вод, второй – из фекалий пациента. Вирусная РНК обнаружена методом РТ-ПЦР с использованием праймеров, специфичных для некодирующих 5' участков энтеровирусов. Результаты. Мы наблюдали присутствие РНК вируса в головном мозге и сердце мышей, инфицированных изолятом из фекалий пациента на 45-й день после заражения (PI). Выводы. Сделан вывод о том, что ВКВ5 сохраняется в мозге и сердце после перорального заражения CD1 мышей. Степень вирусной устойчивости может быть связана с происхождением вируса и его генетикой. Ключевые слова: вирус Коксаки B5, мыши, мозг, устойчивость. 2011 Article Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 / Stipalova D., Sojka M., Borsanyiova M., Badurova M., Marosova L., Sobotova Z., Bopegamage S. // Вiopolymers and Cell. — 2011. — Т. 27, № 2. — С. 162-164. — Бібліогр.: 15 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000091 http://dspace.nbuv.gov.ua/handle/123456789/153709 57.017.2 + 578.835.1 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 |
Short Communications Short Communications |
| spellingShingle |
Short Communications Short Communications Stipalova, D. Sojka, M. Borsanyiova, M. Badurova, M. Marosova, L. Sobotova, Z. Bopegamage, S. Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 Вiopolymers and Cell |
| description |
Aim. The aim of our study was to follow the persistence of viral RNA in selected organs of experimentally
infected with coxsackievirus (CV) B5 strains from different sources such as a patient’s sample, an environmental sample and a prototype virus strain. Methods. CD-1 mice were infected with CVB5 strain
Faulkner the prototype, CVB5 – isolate from treated sewage waste and isolate from patient’s stool sample
both identified as CVB5. The viral RNA was detected by RT-PCR using enterovirus primers specific for the
non-coding 5' region. Results. We observed presence of RNA in the brain and heart of mice infected with
isolate from patient’s stool at day 45 post infection (p. i.). Conclusion. We conclude that CVB5 persists in
the brain and heart after oral infection of CD1 mice. The relevance of viral persistence maybe related viral
origin and the genetics.
Keywords: coxsackievirus B5, mouse, brain, persistence. |
| format |
Article |
| author |
Stipalova, D. Sojka, M. Borsanyiova, M. Badurova, M. Marosova, L. Sobotova, Z. Bopegamage, S. |
| author_facet |
Stipalova, D. Sojka, M. Borsanyiova, M. Badurova, M. Marosova, L. Sobotova, Z. Bopegamage, S. |
| author_sort |
Stipalova, D. |
| title |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 |
| title_short |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 |
| title_full |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 |
| title_fullStr |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 |
| title_full_unstemmed |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 |
| title_sort |
persistence of viral rna in the brain of experimentally infected mice with coxsackievirus b5 |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2011 |
| topic_facet |
Short Communications |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/153709 |
| citation_txt |
Persistence of viral RNA in the brain of experimentally infected mice with coxsackievirus B5 / Stipalova D., Sojka M., Borsanyiova M., Badurova M., Marosova L., Sobotova Z., Bopegamage S. // Вiopolymers and Cell. — 2011. — Т. 27, № 2. — С. 162-164. — Бібліогр.: 15 назв. — англ. |
| series |
Вiopolymers and Cell |
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2025-07-14T05:12:07Z |
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2025-07-14T05:12:07Z |
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| fulltext |
Persistence of viral RNA in the brain of experimentally
infected mice with coxsackievirus B5
D. Stipalova1, M. Sojka1, M. Borsanyiova1, M. Badurova1,
L. Marosova1, Z. Sobotova2, S. Bopegamage1
1Department of Virology, Slovak Medical University
12, Limbova St., Bratislava, Slovak Republic, 83303
2National Reference Centres, Public Health Office of the Slovak Republic
52, Trnavska St., Bratislava, Slovak Republic, 82645
darina.stipalova@szu.sk
Aim. The aim of our study was to follow the persistence of viral RNA in selected organs of experimentally
infected with coxsackievirus (CV) B5 strains from different sources such as a patient’s sample, an en-
vironmental sample and a prototype virus strain. Methods. CD-1 mice were infected with CVB5 strain
Faulkner the prototype, CVB5 – isolate from treated sewage waste and isolate from patient’s stool sample
both identified as CVB5. The viral RNA was detected by RT-PCR using enterovirus primers specific for the
non-coding 5' region. Results. We observed presence of RNA in the brain and heart of mice infected with
isolate from patient’s stool at day 45 post infection (p. i.). Conclusion. We conclude that CVB5 persists in
the brain and heart after oral infection of CD1 mice. The relevance of viral persistence maybe related viral
origin and the genetics.
Keywords: coxsackievirus B5, mouse, brain, persistence.
Introduction. Coxsackievirus B5 (CVB5) belongs to
the genus Enterovirus and the family Picornaviridae.
Epidemiological data have shown that this serotype is
one of the most frequently isolated enteroviruses from
infected humans [1].
CVB5 is associated often with meningitis, sporadic
cases of neurological diseases, and chronic diseases
such as cardiomyopathy and diabetes. Coxsackievirus
and echovirus are frequent causes of aseptic meningitis,
particularly in children, but are rarely life threatening.
Fatality has been reported as in a case of meningoence-
phalitis [2], in immunosuppressed patient.
These viruses exist as circulating heterogeneous
virus populations of genetic variants as other entero-
viruses. Mice are used for studying pathogenesis of
coxsackieviruses due to the presence of mouse coxsa-
ckie-adenovirus receptor (mCAR) which exists on cells
of different organs.
Our aim was to study the persistence of viral RNA
in different organs of experimentally infected mice.
Materials and methods. Virus and cells. CVB5
(Faulkner) was obtained from the former National
Institute of Health, Prague, Czech Republic, and pro-
pagated in Green monkey kidney (GMK) cells. S1 B5
(identified as CVB5) – isolate from the interphase
formed during sewage treatment and S2 B5 (identified
as CVB5) – isolate from patient’s stool sample. GMK
cells were used for virus propagation and titrations.
Cells were grown in Eagle’s minimum essential me-
dium (MEM(E)) supplemented with 10% heat in-
activated bovine serum (at 56 °C) for cell growth and 5
or 2 % serum for maintenance and infection. For plaque
purification the double layer technique was used. The
method was standardized in our laboratory, details have
been described by Motusova et al. [3].
Selected virus plaques were picked up and suspen-
ded in 0.5 ml MEM(E) and used for further stock prepa-
rations and studies.
Virus isolations from organs and stools were
checked as described previously [4] in Hep2 cells.
Mice. CD1 3–4 weeks old (10–12 g) were acquired
from HARLAN (Italy). Mice were housed two or three
162
ISSN 0233–7657. Biopolymers and Cell. 2011. Vol. 27. N 2. P. 162–164
Institute of Molecular Biology and Genetics NAS of Ukraine, 2011
per cage and supplied with sterile water and commer-
cial food pellets (Topdovo). Permission for the animal
work was obtained from the Ethics Committee of the
Slovak Health University and the State Veterinary and
Food Control Authority of the Slovak Republic.
Oral infection. Oral infection of mice has been de-
scribed previously [4, 5]. Mice were divided into four
groups of 20 mice. Each group received 0.5 ml virus
suspension or PBS (uninfected controls). The virus do-
se used for inoculation was 107 TCID50 of each CVB5
(Faulkner), S1 B5 CVB5 – isolate from the interphase
of treated sewage sample and S2 B5 CVB5 – isolate
from patient’s stool sample. Mice were sacrificed and
organs (heart, pancreas, brain) were collected from
each group of 5 infected mice/group and 5 control mice/
group at days 5, 10 and 45 post infection. Organs were
collected separately; snap frozen and stored at –80 oC.
PCR analysis. RNA was extracted from snap-fro-
zen organs (heart, brain, pancreas) with a Mammalian
Total RNA kit («Invitrogen», USA). cDNA synthesis
and cDNA amplification were performed by using
a single tube method with The SuperScript III One-Step
RT-PCR System with Platinum Taq High Fidelity («In-
vitrogen») as has been described [4, 6]. Nested RT-
PCR was done. Primers used were targeted to the 5'
non-coding region as described by de Leeuw et al. [6]
and Bopegamage et al. [4].
Results and discussion. In the present study we ha-
ve studied the presence of viral RNA in organs of CD1
outbred mice infected by three different CVB5 strains
(patient’s isolate and an isolate identified as CVB5
from treated sewage sample and the prototype strain).
Viral RNA was detected in the brain and heart of mice
infected with isolate from patient’s stool at day 45 p. i.
(Table), replicating virus was not isolated at this period
in any of the organs at this time period. The latest
interval studied by us previous to the day 45 p. i. was
day 10 p. i. when the replicating virus is usually iso-
lated. At day 10 p. i. maximum positives were found in
mice infected with the isolate from patient’s stool S2
B5 CVB5. Presence of viral RNA in the brain in an
experimental model has not been studied previously. A
single positive mouse for viral RNA can be explained
due to the outbred mice used by us. Prolonged presence
of viral RNA in a single mouse in the late phases after
infection is common.
In our previous study [4], we had shown prolonged
presence of viral RNA in the pancreas and small in-
testine of Swiss outbred mice infected orally by CVB3
(Nancy strain) given at different doses. CVB infection
and persistence of viral RNA leading to direct damage
of the cardiomyocytes have been studied by Andreoletti
et al. [7], Kandolf et al. [8], and Klingel et al. [9] who
have used inbred A/J mice. CVB3 persistence and in-
bred mouse strains has been reviewed by Chapman and
Kim [10] and Klingel [11]. Persistence of the viral RNA
in the brain of experimentally infected mice by any
CVB has not been studied before. In the last few years
in humans the persistent enteroviruses have been sus-
pected to be the cause of various central nervous system
(CNS) and muscle disorders of unknown etiology, in-
cluding motor neurone disease, post-polio syndrome,
the chronic fatigue syndrome [12]. The mechanism of
viral persistence for enterovirus is not known. The sug-
gested mechanisms include mutations from lytic to
non-lytic or defective mutants and change in cell tro-
pism [13], and the asymmetrical ratio of plus to minus
strand RNA by Tracy [14]. Furthermore mutations in
the stem loop II of the 5' non coding structure has been
suggested by Dunn et al. [15]. Our observations could
be explained as to the difference in the virus strains,
even within the 5 mice in that particular group to the
outbred model used by us. Sequencing of the persistent
RNA from the organs and comparison to the original
viral RNA is required to be studied further.
Conclusion. We conclude that CVB5 persists in the
brain and heart after oral infection of CD1 mice. The
relevance of viral persistence maybe related viral origin
and the genetics.
Acknowledgements. This work was supported by
the Norwegian Financial Mechanism, Mechanism EEA
and Slovak Government and the State Budget of the
Slovak Republic (SK 0082).
163
PROLONGED PRESENCE OF RNA IN THE BRAIN IN MURINE EXPERIMENTAL INFECTION BY COXSACKIEVIRUS B5
Virus Heart Pancreas Brain
CVB5* 0/5 0/5 0/5
S1 B5 0/5 0/5 0/5
S2 B5 1/5** 0/5 1/5
CVB5* – Coxsackievirus B5 strain Faulkner (prototype virus); S1
B5 – isolate from treated sewage sample identified as CVB5 labeled as
S1; S2 B5 – isolate from patient’s stool identified as CVB5 labeled as
S2; **number of organs positive for viral RNA/total number of organs
tested.
Presence of viral RNA in organs of orally infected CD1 outbred
mice at day 45 p. i.
Д. Стіпа ло ва, М. Сой ка, М. Бор сань йо ва, М. Ба ду ро ва,
Л. Ма ро зо ва, З. Шо бо то ва, С. Бо пе га ма ге
Збе ре жен ня вірус ної РНК у го лов но му моз ку ми шей,
ек спе ри мен таль но інфіко ва них віру сом Кок сакі В5
Ре зю ме
Мета на шо го досліджен ня по ля га ла у тому, щоб про слідку ва -
ти, як вірус на РНК зберігається в окре мих орга нах ми шей, ек-
спе ри мен таль но інфіко ва них де я ки ми шта ма ми вірусу Кок сакі
(ВК) B5, виділе но му з різних дже рел: із зраз ка пацієнта, на вко -
лиш ньо го се ре до ви ща і шта му про то ти пу вірусу. Ме то ди. Ми -
шей CD-1 інфіку ва ли ВКВ5 – про то ти пом Фол кне ра і дво ма
віру са ми, іден тифіко ва ни ми як ВКВ5: пер ший виділено з об -
роб ле них відходів стічних вод, дру гий – із фе калій пацієнта.
Вірус ну РНК ви яв ле но ме то дом РТ-ПЛР з ви ко рис тан ням
прай мерів, спе ци фич них для не ко ду ю чих 5'-діля нок ен те рові-
русів. Ре зуль та ты. Ми спос терігали при сутність РНК вірусу
в го лов но му моз ку і серці ми шей, інфіко ва них ізо ля том із фека-
лій пацієнта на 45-й день після за ра жен ня (PI). Вис нов ки.
Зроб ле но вис но вок сто сов но того, що ВКВ5 зберігається в
моз ку і серці після пе ро раль но го за ра жен ня CD1 ми шей. Сту-
пінь вірус ної стійкості можу бути по в’я за ний з по ход жен ням
вірусу та його ге не ти кою.
Клю чові сло ва: вірус Кок сакі B5, миші, мо зок, стійкість.
Д. Сти па ло ва, М. Сой ка, М. Бор сань йо ва, М. Ба ду ро ва,
Л. Ма ро зо ва, З. Шо бо то ва, С. Бо пе га ма ге
Сох ра не ние ви рус ной РНК в го лов ном моз ге мы шей,
экс пе ри мен таль но ин фи ци ро ван ных ви ру сом Кок са ки В5
Ре зю ме
Цель на ше го ис сле до ва ния со сто я ла в том, что бы про сле -
дить, как ви рус ная РНК со хра ня ет ся в от дель ных орга нах мы -
шей, экс пе ри мен таль но ин фи ци ро ван ных раз лич ны ми штам-
мами ви ру са Кок са ки (ВК) B5, вы де лен ны ми из раз ных ис точ -
ни ков: из об раз ца па ци ен та, окру жа ю щей сре ды и штам ма
про то ти па ви ру са. Ме то ды. Мы шей CD-1 ин фи ци ро ва ли
ВКВ5 – про то ти пом Фол кне ра и дву мя ви ру са ми, иден ти фи ци -
ро ван ны ми как ВКВ5: пер вый вы де лен из об ра бо тан ных от хо -
дов сточ ных вод, вто рой – из фе ка лий па ци ен та. Ви рус ная
РНК об на ру же на ме то дом РТ-ПЦР с ис поль зо ва ни ем прай ме -
ров, спе ци фич ных для неко ди ру ю щих 5' учас тков эн те ро ви ру -
сов. Ре зуль та ты. Мы на блю да ли при су тствие РНК ви ру са в
го лов ном моз ге и сер дце мы шей, ин фи ци ро ван ных изо ля том из
фе ка лий па ци ен та на 45-й день по сле за ра же ния (PI). Вы во ды.
Сде лан вы вод о том, что ВКВ5 со хра ня ет ся в моз ге и сер дце
по сле пе ро раль но го за ра же ния CD1 мы шей. Сте пень ви рус ной
устой чи вос ти мо жет быть свя за на с про ис хож де ни ем ви ру са
и его ге не ти кой.
Клю че вые сло ва: ви рус Кок са ки B5, мыши, мозг, устой чи вость.
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UDC 57.017.2 + 578.835.1
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