Visual and rapid detection of Plesiomonas shigelloides using loop‐mediated isothermal amplification method

2019 ◽  
Vol 69 (6) ◽  
pp. 411-416
Author(s):  
H. Liu ◽  
H. Yu ◽  
Z.Q. Peng ◽  
Y.Y. Yu ◽  
J.F. Xie ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Isothermal Amplification ◽  
Plesiomonas Shigelloides ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method

scholarly journals Sensitive and Rapid Detection of Edwardsiellosis in Fish by a Loop-Mediated Isothermal Amplification Method

2004 ◽  
Vol 70 (1) ◽  
pp. 621-624 ◽  
Author(s):  
Ram Savan ◽  
Arisa Igarashi ◽  
Satoru Matsuoka ◽  
Masahiro Sakai
Keyword(s):  
Rapid Detection ◽  
Japanese Flounder ◽  
Isothermal Amplification ◽  
Edwardsiella Tarda ◽  
Fish Disease ◽  
Fish Pathogen ◽  
Loop Mediated Isothermal Amplification ◽  
Diagnostic Protocol ◽  
Amplification Method ◽  
Hemolysin Gene

ABSTRACT Here we report a rapid and sensitive method (using loop-mediated isothermal amplification [LAMP]) for the diagnosis of edwardsiellosis, a fish disease caused by Edwardsiella tarda, in Japanese flounder. A set of four primers was designed, and conditions for the detection were optimized for the detection of E. tarda in 45 min at 65°C. No amplification of the target hemolysin gene was detected in other related bacteria. When the LAMP primers were used, detection of edwardsiellosis in infected Japanese flounder kidney, and spleen and seawater cultures was possible. We have developed a rapid and sensitive diagnostic protocol for edwardsiellosis detection in fish. This is the first report of the application of LAMP for the diagnosis of a fish pathogen.

scholarly journals Development of a Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Phytopythium vexans

2021 ◽  
Vol 12 ◽  
Author(s):  
Tuhong Wang ◽  
Haojun Ji ◽  
Yongting Yu ◽  
Xiaojie Wang ◽  
Yi Cheng ◽  
...  
Keyword(s):  
Root Rot ◽  
Rapid Detection ◽  
Color Change ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method ◽  
Specialized Equipment ◽  
Root Tissues ◽  
And Control ◽  
Brown Root Rot

Brown root rot caused by Phytopythium vexans is a new destructive root disease on many plants such as Gingko, Citrus, kiwifruit, and ramie. The establishment of loop-mediated isothermal amplification (LAMP) technology for detecting P. vexans can help monitor and control brown root rot quickly, efficiently, and accurately. LAMP technology is known for its simplicity, sensitivity, and speed; and it does not require any specialized equipment – a water bath or a thermoblock is sufficient for isothermal amplifications. LAMP products can be visualized by using hydroxy naphthol blue (HNB) dye or agarose gel electrophoresis. In this study, by searching and comparing the internal transcribed spacer (ITS) sequences of P. vexans and the related species in oomycete genera Pythium, Phytopythium, and Phytophthora, we designed specific primers targeting the ITS gene region of P. vexans. Using HNB dye, we established a LAMP technique for rapid detection of P. vexans by visible color change. In addition, we optimized the protocol to enhance both sensitivity and specificity for P. vexans detection. Under the optimized condition, our protocol based on LAMP technology could detect as low as 24 copies of the P. vexans genomic DNA, which is ∼100 times more sensitive than conventional PCR. This method can successfully detect P. vexans using cell suspensions from P. vexans – infected ramie root tissues.

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