Multiplex polymerase chain reaction (PCR) with Nanopore sequencing for sequence-based detection of four tilapia pathogens DOI Creative Commons
Jérôme Delamare‐Deboutteville,

Watcharachai Meemetta,

Khaettareeya Pimsannil

et al.

PeerJ, Journal Year: 2025, Volume and Issue: 13, P. e19425 - e19425

Published: May 13, 2025

Tilapia aquaculture faces significant threats posed by four prominent pathogens: tilapia lake virus (TiLV), infectious spleen and kidney necrosis (ISKNV), Francisella orientalis, Streptococcus agalactiae. Currently, employed molecular diagnostic methods for these pathogens rely on multiple singleplex polymerase chain reactions (PCR), which are time-consuming expensive. In this study, we present an approach utilizing a multiplex PCR (mPCR) assay, coupled with rapid Nanopore sequencing, enabling the one-tube simultaneous detection one-reaction sequencing-based validation of pathogens. Our assay exhibits limit 1,000 copies per reaction TiLV, ISKNV, S. agalactiae, while F. is 10,000 reaction. This sensitivity sufficient diagnosing infections co-infections in clinical samples from sick fish, confirmation presence Integrating sequencing provides alternative platform fast precise diagnostics major clinically animals, adding to available toolbox disease diagnostics.

Language: Английский

Multiplex polymerase chain reaction (PCR) with Nanopore sequencing for sequence-based detection of four tilapia pathogens DOI Creative Commons
Jérôme Delamare‐Deboutteville,

Watcharachai Meemetta,

Khaettareeya Pimsannil

et al.

PeerJ, Journal Year: 2025, Volume and Issue: 13, P. e19425 - e19425

Published: May 13, 2025

Tilapia aquaculture faces significant threats posed by four prominent pathogens: tilapia lake virus (TiLV), infectious spleen and kidney necrosis (ISKNV), Francisella orientalis, Streptococcus agalactiae. Currently, employed molecular diagnostic methods for these pathogens rely on multiple singleplex polymerase chain reactions (PCR), which are time-consuming expensive. In this study, we present an approach utilizing a multiplex PCR (mPCR) assay, coupled with rapid Nanopore sequencing, enabling the one-tube simultaneous detection one-reaction sequencing-based validation of pathogens. Our assay exhibits limit 1,000 copies per reaction TiLV, ISKNV, S. agalactiae, while F. is 10,000 reaction. This sensitivity sufficient diagnosing infections co-infections in clinical samples from sick fish, confirmation presence Integrating sequencing provides alternative platform fast precise diagnostics major clinically animals, adding to available toolbox disease diagnostics.

Language: Английский

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