NDLI: Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

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Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

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Acknowledgment

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Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

Content Provider	Springer Nature Link
Author	Fall, Jean Chakraborty, Gunimala Kono, Tomoya Maeda, Minoru Suzuki, Yoshihiro Itami, Toshiaki Sakai, Masahiro
Copyright Year	2010
Abstract	Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10$^{2}$ copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T$_{t}$) versus the log of bacterial number. A high correlation coefficient (R$^{2}$ = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.
Starting Page	129
Ending Page	134
Page Count	6
File Format	PDF
ISSN	09199268
Journal	Fisheries Science
Volume Number	77
Issue Number	1
e-ISSN	14442906
Language	English
Publisher	Springer Japan
Publisher Date	2010-12-07
Publisher Place	Japan
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Quantitative loop-mediated isothermal amplification Vibrio nigripulchritudo Intergenic spacer region DNA Food Science Freshwater & Marine Ecology Fish & Wildlife Biology & Management
Content Type	Text
Resource Type	Article
Subject	Aquatic Science

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