Rapid and sensitive detection ofVibrio harveyiby loop-mediated isothermal amplification combined with lateral flow dipstick targeted tovhhP2gene

Rapid and Sensitive Detection of Vibrio alginolyticus by Loop‐Mediated Isothermal Amplification Combined with a Lateral Flow Dipstick Targeted to the rpoX Gene

“…In pure culture, the detection limit of LAMP-LFD was 1.8 × 10 2 CFU/mL (0.6 CFU per reaction, which is one-tenth that of PCR). The results of the sensitivity study were similar to those of earlier LAMP-LFD assays for V. parahaemolyticus (0.4 CFU per reaction; Prompamorn et al 2011) and V. harveyi (0.6 CFU per reaction; Thongkao et al 2015) and a typical LAMP assay targeting the gyrB gene of V. alginolyticus (3.7 × 10 2 CFU/mL [3.7 CFU per reaction]; Cai et al 2010). However, the higher detection limit at 0.6 CFU/reaction may relate to DNA from nonviable bacterial cells or viable but nonculturable cells (Du et al 2007), which could have influenced the detection limit.…”

“…The LFD can be immersed into a buffer solution containing LAMP amplicons and the color can be readily observed. The LAMP-LFD method has been utilized to identify bacterial pathogens in marine animal culture, such as V. vulnificus (Surasilp et al 2011), V. parahaemolyticus (Prompamorn et al 2011), and V. harveyi (Thongkao et al 2015). In the present study, a LAMP-LFD assay targeting the rpoX gene of V. alginolyticus was developed.…”

Development of Duplex Loop‐Mediated Isothermal Amplification (dLAMP) Combined with Lateral Flow Dipstick (LFD) for the Rapid and Specific Detection of Vibrio vulnificus and V. parahaemolyticus

“…In most cases, the sensitivity of a monoplex LAMP assay, such as LAMP-LFD for V. harveyi detection in added shrimp samples (5 CFU per reaction: Thongkao et al 2015), conventional LAMP for V. harveyi in spiked shellfish cultures (17.2 CFU per reaction: Cao et al 2010), and V. cholera in added shrimp samples (20 CFU per reaction: Srisuk et al 2010) was greater than that of dLAMP-LFD in our study. The lower sensitivity may be due to competition among the number of primers that were used under multidetection conditions, as previously described (Ali and Reynolds 2000;Pang et al 2002;Xie et al 2006;Rashid et al 2009).…”

“…This approach not only avoids using carcinogens, such as ethidium bromide, but also improves the sensitivity of the assay (Wang et al 2012). The LAMP-LFD assay has been used to verify the viral pathogens in shrimp culture, such as hepatopancreatic parvovirus (Nimitphak et al 2008), white spot syndrome virus (Jaroenram et al 2009), infectious myonecrosis virus (Puthawibool et al 2009), and Macrobrachium rosenbergii nodavirus (Puthawibool et al 2010), and bacterial pathogens, such as V. vulnificus (Surasilp et al 2011), V. parahaemolyticus (Prompamorn et al 2011), and V. harveyi (Thongkao et al 2015).…”

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

“…The sample of interest, for example, saliva, blood, urine and other body fluids, is applied at one end of the strip (sample pad), which is conjugated with LAMP buffer mix and biotin, making the loaded sample suitable for interaction with the detection system. Most generally, lateral flow test was established using fluorescein in isothiocyanate (FITC) labelled DNA probes (which will recognize the specific region in the LAMP amplicon) hybridized with biotinylated LAMP amplicons (Thongkao et al 2015;Fowler et al 2016;Huang et al 2017;Park et al 2017). This complex would be captured by streptavidin on the biotin, forming another complex with anti-FITC antibody coated on the gold nanoparticles.…”