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Development of a novel multiplex-PCR technology for simultaneous detection of five major aquaculture pathogens

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Title Development of a novel multiplex-PCR technology for simultaneous detection of five major aquaculture pathogens
 
Creator Nair, Anusree V.
Sumithra, TG
Reshma, KJ
Sanil, NK
 
Subject Internal amplification control
Vibrio spp
 
Description 379-388
Early and precise pathogen identification and corresponding disease management are primary concerns in aquaculture.
Here, we attempted at diagnostic methods that can simultaneously identify multiple pathogens, where many samples, several
pathogens, and concurrent infections are to be handled. Hence, a multiplex PCR assay targeting five major aquaculture
pathogens, viz. Vibrio parahaemolyticus, Vibrio anguillarum, Vibrio alginolyticus, Vibrio vulnificus and Vibrio harveyi was
developed for the first time. The primers targeting toxR of V. parahaemolyticus, amiB of V. anguillarum, col of
V. alginolyticus, vvhA of V. vulnificus, and topA of V. harveyi were applied. Furthermore, the reaction included an internal
amplification control against prokaryotic 16S rRNA to perceive false-negative results. The assay showed 100% specificity
against 56 unique bacteria. The sensitivity was 0.25 ng for V. harveyi, 0.5 ng for V. vulnificus, 1 ng for V. parahaemolyticus
and V. anguillarum, and 2 ng for V. alginolyticus DNA per μL assay. Sensitivity regarding CFU was 1.2, 5.2, 10, 5.6×101
and 3.8×102 per μL, for V. harveyi, V. vulnificus, V. anguillarum, V. parahaemolyticus and V. alginolyticus, respectively.
The results suggest that the optimized method can be applied for sensitive and specific identification of five aquaculture
pathogens through a single PCR.
 
Date 2023-05-01T06:57:17Z
2023-05-01T06:57:17Z
2023-05
 
Type Article
 
Identifier 0975-1009 (Online); 0019-5189 (Print)
http://nopr.niscpr.res.in/handle/123456789/61823
https://doi.org/10.56042/ijeb.v61i05.857
 
Language en
 
Publisher NIScPR-CSIR, India
 
Source IJEB Vol.61(05) [May 2023]