Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi
Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this stud...
| Autor principal: | ALMEIDA, Sintia Silva de |
|---|---|
| Outros Autores: | DORNELES, Elaine Maria Seles, DINIZ, Carlos Augusto Almeida, ABREU, Vinicius Augusto Carvalho de, SOUSA, Cassiana Severiano de, ALVES, Jorianne Thyeska Castro, CARNEIRO, Adriana Ribeiro, VILAS BOAS, Priscilla Carolinne Bagano, SPIER, Sharon, BARH, Debmalya, LAGE, Andrey Pereira, FIGUEIREDO, Henrique Cesar Pereira, AZEVEDO, Vasco Ariston de Carvalho |
| Grau: | Artigo |
| Idioma: | eng |
| Publicado em: |
BioMed Central Ltd
2023
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| Assuntos: | |
| Acesso em linha: |
https://repositorio.ufpa.br/jspui/handle/2011/15726 |
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ir-2011-15726 |
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ir-2011-157262023-06-21T17:44:52Z Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi ALMEIDA, Sintia Silva de DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis. Methods: In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the diagnosis of the multiplex PCR at biovar level. Results: A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed. The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type strain ATCC 19410T , were compared to the results of nitrate reductase identification by biochemical test. The McNemar’s Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16–6.14)] between the quadruplex PCR and the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and the kappa value was 0.94 [95% CI (0.90–0.98)]. Conclusions: The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and Equi strains enhances its usefulness in the clinical microbiology laboratory. CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais ALVES, J. T. C.; CARNEIRO, A. R. Universidade Federal do Pará 2023-06-21T17:42:35Z 2023-06-21T17:42:35Z 2017 Artigo de Periódico ALMEIDA, Sintia et al. Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi. BMC Genomics, online, v. 13, n. 390, p. 1-8, 2017. DOI: https://doi.org/10.1186/s12917-017-1210-5. Disponível em: https://repositorio.ufpa.br/jspui/handle/2011/15726. Acesso em:. 1471-2164 https://repositorio.ufpa.br/jspui/handle/2011/15726 10.1186/s12917-017-1210-5 eng BMC Genomics Acesso Aberto Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by/4.0/ application/pdf BioMed Central Ltd Reino Unido https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-017-1210-5#Ack1 |
| institution |
Repositório Institucional - Universidade Federal do Pará |
| collection |
RI-UFPA |
| language |
eng |
| topic |
Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats |
| spellingShingle |
Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats ALMEIDA, Sintia Silva de Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| topic_facet |
Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats |
| description |
Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which
is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes
abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay
that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis.
Methods: In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved
in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate
reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the
diagnosis of the multiplex PCR at biovar level.
Results: A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed.
The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis
from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type
strain ATCC 19410T
, were compared to the results of nitrate reductase identification by biochemical test. The
McNemar’s Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification
showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16–6.14)] between the quadruplex PCR and
the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and
the kappa value was 0.94 [95% CI (0.90–0.98)].
Conclusions: The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and
Equi strains enhances its usefulness in the clinical microbiology laboratory. |
| format |
Artigo |
| author |
ALMEIDA, Sintia Silva de |
| author2 |
DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho |
| author2Str |
DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho |
| title |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| title_short |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| title_full |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| title_fullStr |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| title_full_unstemmed |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
| title_sort |
quadruplex pcr assay for identification of corynebacterium pseudotuberculosis differentiating biovar ovis and equi |
| publisher |
BioMed Central Ltd |
| publishDate |
2023 |
| url |
https://repositorio.ufpa.br/jspui/handle/2011/15726 |
| _version_ |
1787147998630772736 |
| score |
11.675088 |