Extension of multiplex PCR for Streptococcus suis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F24%3AN0000104" target="_blank" >RIV/00027162:_____/24:N0000104 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.eavld2024.org/wp-content/uploads/2024/10/Programma-DEFINITIVO-WEB.pdf" target="_blank" >https://www.eavld2024.org/wp-content/uploads/2024/10/Programma-DEFINITIVO-WEB.pdf</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Extension of multiplex PCR for Streptococcus suis

Popis výsledku v původním jazyce

Introduction: Streptococcus suis is an important zoonotic pathogen of swine. 29 different capsular serotypes have been described and novel cps loci are constantly emerging [1-5]. To distinguish between serotypes, Multiplex PCR tests have recently been developed that target specific genes in the cps loci of S. suis [6]. Here, we have developed an extension of the multiplex PCR method from Kerdsin et al. for non-serotypeable strains collected in the Czech Republic. Materials and Methods: After Whole genome sequencing of S. suis isolates collected in the Czech Republic and analyzing the cps loci of non-serotypeable isolates, we designed 19 pairs of specific primers targeting unique sequences to detect potentially novel cps loci. After verifying the functionality of Singleplex PCR reactions, we developed three Multiplex PCRs to detect multiple isolates simultaneously. Results: Initially, we successfully tested all 19 PCR reactions. We developed Multiplex PCR schemes for 15 primers, while the remaining 4 primers, with similar PCR product lengths, have not yet been included in the Multiplex PCR. We successfully tested two Multiplex PCRs, each capable of distinguishing five types. Despite successful Singleplex PCRs, our attempt to verify the functionality of five additional primer pairs in the third Multiplex PCR was unsuccessful, indicating the need for optimization of the reaction conditions. Discussion and Conclusion: We successfully developed two Multiplex PCRs, which can distinguish a total of 10 possible strains in two reactions. Although we were unable to design a third Multiplex PCR for the simultaneous differentiation of five strains, we believe that optimizing the conditions will allow us to achieve this. The extended Multiplex PCR allows for the identification of additional serotypes, thus reducing the number of untypeable S. suis strains. This work was supported by grants RO0523 and TN02000017.

Název v anglickém jazyce

Extension of multiplex PCR for Streptococcus suis

Popis výsledku anglicky

Introduction: Streptococcus suis is an important zoonotic pathogen of swine. 29 different capsular serotypes have been described and novel cps loci are constantly emerging [1-5]. To distinguish between serotypes, Multiplex PCR tests have recently been developed that target specific genes in the cps loci of S. suis [6]. Here, we have developed an extension of the multiplex PCR method from Kerdsin et al. for non-serotypeable strains collected in the Czech Republic. Materials and Methods: After Whole genome sequencing of S. suis isolates collected in the Czech Republic and analyzing the cps loci of non-serotypeable isolates, we designed 19 pairs of specific primers targeting unique sequences to detect potentially novel cps loci. After verifying the functionality of Singleplex PCR reactions, we developed three Multiplex PCRs to detect multiple isolates simultaneously. Results: Initially, we successfully tested all 19 PCR reactions. We developed Multiplex PCR schemes for 15 primers, while the remaining 4 primers, with similar PCR product lengths, have not yet been included in the Multiplex PCR. We successfully tested two Multiplex PCRs, each capable of distinguishing five types. Despite successful Singleplex PCRs, our attempt to verify the functionality of five additional primer pairs in the third Multiplex PCR was unsuccessful, indicating the need for optimization of the reaction conditions. Discussion and Conclusion: We successfully developed two Multiplex PCRs, which can distinguish a total of 10 possible strains in two reactions. Although we were unable to design a third Multiplex PCR for the simultaneous differentiation of five strains, we believe that optimizing the conditions will allow us to achieve this. The extended Multiplex PCR allows for the identification of additional serotypes, thus reducing the number of untypeable S. suis strains. This work was supported by grants RO0523 and TN02000017.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

40301 - Veterinary science

Projekt

<a href="/cs/project/TN02000017" target="_blank" >TN02000017: Národní Centrum Biotechnologií ve Veterinární Medicíně</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů