Detalhes bibliográficos
| Ano de defesa: |
2019 |
| Autor(a) principal: |
CORTEZ, ANNIRA AQUINO |
| Orientador(a): |
Não Informado pela instituição |
| Banca de defesa: |
Não Informado pela instituição |
| Tipo de documento: |
Tese
|
| Tipo de acesso: |
Acesso aberto |
| Idioma: |
por |
| Instituição de defesa: |
Universidade Estadual do Ceará
|
| Programa de Pós-Graduação: |
Não Informado pela instituição
|
| Departamento: |
Não Informado pela instituição
|
| País: |
Não Informado pela instituição
|
| Palavras-chave em Português: |
|
| Link de acesso: |
https://siduece.uece.br/siduece/trabalhoAcademicoPublico.jsf?id=90453
|
Resumo: |
<!--[if !mso]> <style> v\:* {behavior:url(#default#VML);} o\:* {behavior:url(#default#VML);} w\:* {behavior:url(#default#VML);} .shape {behavior:url(#default#VML);} </style> <![endif]--> <div class="WordSection1"> <p class="MsoBodyText" style="margin-top:0cm;margin-right:7.35pt;margin-bottom: 0cm;margin-left:17.1pt;margin-bottom:.0001pt;text-align:justify;line-height: 150%">O objetivo desse trabalho foi desenvolver uma técnica diagnóstica de PCR Multiplex eficaz na identificação de cepas <span style="letter-spacing:.1pt">de </span><i style="mso-bidi-font-style:normal">Brucella </i>sp<i style="mso-bidi-font-style:normal">. </i>em suínos, com a finalidade de implantação na vigilância da Brucelose suína na defesa agropecuária. O trabalho foi dividido em dois experimentos: no primeiro, utilizaram-se quatorze amostras de sêmen e sangue total de varrões e no segundo, foram coletadas 109 amostras de sangue total de suínos abatidos em um abatedouro na região metropolitana de Fortaleza. Das amostras de sangue, foi extraído soro sanguíneo, que foi submetido ao teste de Antígeno <span style="letter-spacing:-.2pt">Tamponado </span>Acidificado <span style="letter-spacing:-.2pt">(AAT) </span>e caso reagente, submetido ao teste de Soroaglutinação Lenta (SAL) e teste de 2-Mercaptoetanol (2-ME). Para o PCR Multiplex, foram utilizadas alíquotas de 100 μL e 200 μL de sêmen e sangue total,<span style="mso-spacerun:yes"> </span>que foram<span style="mso-spacerun:yes"> </span>lavadas<span style="mso-spacerun:yes"> </span>quatro<span style="mso-spacerun:yes"> </span>vezes<span style="mso-spacerun:yes"> </span>em<span style="mso-spacerun:yes"> </span>solução<span style="mso-spacerun:yes"> </span>de<span style="mso-spacerun:yes"> </span>lavagem e<span style="letter-spacing:-.05pt"> </span>centrifugadas a</p> <p class="MsoBodyText" style="margin-top:.1pt;margin-right:7.05pt;margin-bottom: 0cm;margin-left:17.1pt;margin-bottom:.0001pt;text-align:justify;line-height: 150%">3.000 g por 1 min, sendo o sedimento final ressuspenso em 60 <span style="letter-spacing:.1pt">µl </span>de solução à base de NaOH, incubado a 65 °C por 10 min e adicionado a 60 <span style="letter-spacing:.1pt">µL </span>de solução base de TrisHCl. Em seguida, foram utilizados 0,5 μL de oito pares de primers de <i style="mso-bidi-font-style:normal">Brucella </i>sp<i style="mso-bidi-font-style:normal">.</i>, 12,5 μL de MasterMix, 2,5 μL de Água MilliQ e 2 μL da amostra, constituindo uma solução de 25 μL, submetidos à amplificação no protocolo de PCR Multiplex. <span style="letter-spacing:-.3pt">Todos </span>os varrões apresentaram <span style="letter-spacing:-.35pt">AAT </span>negativo e 26 suínos abatidos foram <span style="letter-spacing:-.35pt">AAT </span>positivos. <span style="letter-spacing:-.3pt">Todas </span>as amostras de sêmen foram negativas no PCR Multiplex. Seis varrões apresentaram sequência-alvo de 272 pb, compatíveis a <i style="mso-bidi-font-style:normal">B. suis, </i>e um varrão apresentou três sequências-alvo (794, 587 e 450 pb), compatíveis com <i style="mso-bidi-font-style:normal">B. abortus </i>cepa vacinal RB51, nas amostras de 100 μL. Nas amostras de 200 μL, um varrão apresentou sequência-alvo de 272 pb e um varrão apresentou duas sequências- alvo, de 450 e 272 pb. Dos Trinta e seis amostras dos suínos abatidos, submetidos ao PCR Multiplex, dois suínos apresentam amplificação de sequências-alvo de 272 pb nas amostras de 100 μL, dois suínos com três sequências-alvo (794, 587 e 450 pb) nas amostras de 100 μL, um suíno com amplificação de três sequências-alvo (794, 587 e 450 pb) nas amostras de 200 μL e quatro suínos com amplificação de três sequências-alvo (794, 587 e 450 pb) nas amostras de 100 e 200 μL, sendo nove suínos positivos no PCR Multiplex. Assim, foi possível diagnosticar diferentes cepas de<span style="mso-spacerun:yes"> </span><i style="mso-bidi-font-style:normal">Brucella<span style="mso-spacerun:yes"> </span></i>sp.<span style="mso-spacerun:yes"> </span>utilizando<span style="mso-spacerun:yes"> </span>sangue<span style="mso-spacerun:yes"> </span>total<span style="mso-spacerun:yes"> </span>de<span style="mso-spacerun:yes"> </span>varrões<span style="mso-spacerun:yes"> </span>e<span style="mso-spacerun:yes"> </span>suínos<span style="mso-spacerun:yes"> </span>abatidos<span style="mso-spacerun:yes"> </span>no<span style="letter-spacing:2.85pt"> </span>Ceará</p></div><div class="WordSection2"><table cellspacing="0" cellpadding="0" align="left"><tbody><tr><td height="45" width="691"><br/></td></tr><tr><td><br/></td><td><br/></td> </tr> </tbody></table> <span style="mso-ignore:vglayout;position: absolute;z-index:-1;margin-left:706px;margin-top:125px;width:56px;height:43px"><img src="file:///C:\DOCUME~1\LEILA~1.SAT\CONFIG~1\Temp\msohtmlclip1\01\clip_image002.gif" height="43" width="56"/></span><span style="font-size:10.0pt; mso-bidi-font-size:12.0pt"></span> <p class="MsoBodyText"><span style="font-size:10.0pt;mso-bidi-font-size:12.0pt"> </span></p> <p class="MsoBodyText"><span style="font-size:10.0pt;mso-bidi-font-size:12.0pt"> </span></p> <p class="MsoBodyText" style="margin-top:.3pt"><span style="font-size:11.0pt; mso-bidi-font-size:12.0pt"> </span></p> <br style="mso-ignore:vglayout" clear="ALL"/> <p class="MsoBodyText" style="margin-top:4.6pt;margin-right:0cm;margin-bottom: 0cm;margin-left:17.1pt;margin-bottom:.0001pt;line-height:150%"><span style="mso-ignore:vglayout;position: relative;z-index:-3"><span style="left:0px;position:absolute;left:710px; top:-31px;width:12px;height:22px"> </span></span></p><table cellspacing="0" cellpadding="0"> <tbody><tr> <td style="vertical-align:top" height="22" width="12"><span style="position:absolute;mso-ignore:vglayout;left:0pt;z-index:-3"> </span><table cellspacing="0" cellpadding="0" width="100%"> <tbody><tr> <td> <div style="padding:0pt 0pt 0pt 0pt" class="shape"> <p class="MsoBodyText" style="line-height:13.3pt;mso-line-height-rule:exactly"><span style="font-family:"Times New Roman","serif";mso-hansi-font-family:Arial; mso-bidi-font-family:Arial">9</span></p> </div> </td> </tr> </tbody></table> </td> </tr> </tbody></table> <span style="mso-ignore:vglayout;position: absolute;z-index:-2;left:0px;margin-left:705px;margin-top:34px;width:9px; height:22px"> </span><table cellspacing="0" cellpadding="0"> <tbody><tr> <td style="vertical-align:top" height="22" width="9"><span style="position:absolute;mso-ignore:vglayout;left:0pt;z-index:-2"> </span><table cellspacing="0" cellpadding="0" width="100%"> <tbody><tr> <td> <div style="padding:0pt 0pt 0pt 0pt" class="shape"> <p class="MsoBodyText" style="line-height:13.4pt;mso-line-height-rule:exactly">.</p> </div> </td> </tr> </tbody></table> </td> </tr> </tbody></table> através de um protocolo de PCR multiplex. Enquanto que são necessários mais estudos para a aplicação da técnica no diagnóstico de <i style="mso-bidi-font-style:normal">Brucella </i>sp. no sêmen suíno <p class="MsoBodyText" style="margin-top:.55pt"><span style="font-size:17.5pt; mso-bidi-font-size:12.0pt"> </span></p> <p class="MsoBodyText" style="margin-left:17.1pt;line-height:150%"><b style="mso-bidi-font-weight:normal">Palavras-chave: </b>Brucelose. Biologia molecular. Suinocultura. Reprodução. Doença infecciosa. 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