Evaluation of loop-mediated isothermal amplification assay along with conventional and real-time PCR assay for sensitive detection of pathogenic Vibrio parahaemolyticus from seafood sample without enrichment

Anupama, Karanth Padyana; Nayak, Ashwath; Karunasagar, Iddya; Maiti, Biswajit

doi:10.1007/s11033-020-06116-9

Cited by 14 publications

(3 citation statements)

References 36 publications

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“…For example, LAMP usually achieves a higher analytical sensitivity only when compared to conventional PCR 17 , 18 . Considering results of real-time PCR, similar or even worse detection limits are frequently obtained 19 , 20 . Therefore, it was not unusual that the TP LAMP assay presented in this study showed an analytical sensitivity 10 times lower than that of real-time PCR.…”

Section: Discussionmentioning

confidence: 87%

Identification of the novel potential pathogen Trueperella pecoris with interspecies significance by LAMP diagnostics

Kreitlow,

Ningrum,

Lämmler

et al. 2023

Sci Rep

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Trueperella pecoris was described as a new species of the genus Trueperella in 2021 and might be pathogenic to various animal species. However, the lack of a suitable diagnostic test system stands in the way of epidemiological surveys to clarify possible causalities. In this study, a Loop-mediated Isothermal Amplification (LAMP) assay was developed and validated that was highly specific for T. pecoris. The assay provided an analytical sensitivity of 0.5 pg/25 µL and showed 100% inclusivity and exclusivity for 11 target and 33 non-target strains, respectively. Three different DNA extraction methods were evaluated to select the most LAMP-compatible method for cell disruption in pure and complex samples. Using an on-site applicable single-buffer DNA extraction with additional heating, the cell-based detection limit was 2.3 CFU/reaction. Finally, the LAMP assay was validated by means of artificially contaminated porcine lung tissue samples in which minimal microbial loads between 6.54 and 8.37 × 103 CFU per swab sample were detectable. The LAMP assay established in this study represents a suitable diagnostic procedure for identifying T. pecoris in clinical specimens and will help to collect epidemiological data on the pathogenicity of this species.

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Section: Discussionmentioning

confidence: 87%

Identification of the novel potential pathogen Trueperella pecoris with interspecies significance by LAMP diagnostics

Kreitlow,

Ningrum,

Lämmler

et al. 2023

Sci Rep

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show abstract

“…10,13 Furthermore, ASEA in detecting cultured V. parahaemolyticus was even more sensitive (about 10 times higher) in comparison with the detection limit of 3.7 CFU per reaction for LAMP. 32 Polyacrylamide gel electrophoresis (PAGE) showed that the ASEA target band of 67 bp (Fig. 4B) and SEA target band of 45 bp (Fig.…”

Section: Comparison Of Asea and Sea In The Detection Of Pathogenic V ...mentioning

confidence: 99%

Rapid, specific and sensitive detection of Vibrio parahaemolyticus in seafood by accelerated strand exchange amplification

et al. 2023

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“…V. parahaemolyticus is a halophilic pathogen that can be found in marine, estuarine, and aquaculture environments all over the world, and is detected constantly in aquatic products such as shellfish, crustaceans, fish, and cephalopods [7,8]. Gastroenteritis is usually caused by eating contaminated shellfish and other raw or undercooked aquatic products containing V. parahaemolyticus [9]. V. vulnificus is another halophilic pathogen found in coastal areas and estuaries.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Pathogenic Vibrio Contamination in Marine Products along China Based on Fluorescence Quantitative PCR

Wang

Wan

Huang

et al. 2022

Journal of Food Quality

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At present, aquatic product pollution has become the main root of frequent food safety problems and causes economic losses. Vibrio is one of the main pathogens causing foodborne diseases. In this study, in order to uncover the pollution status of pathogenic Vibrio in the marine products of China, a total of 646 aquatic products were collected and analyzed from 10 coastal cities in China. Five kinds of pathogenic Vibrio were separated from these samples and monitored to explore the relationship between pollution and the pathogen. Real-time fluorescence quantitative PCR was adopted to detect foodborne Vibrio quantitatively in marine aquatic products. Aquatic pathogenic Vibrio was collected in different regions, different types of aquatic products, and different sampling places, and the difference in detection rate was statistically significant through statistical analysis. This study made a frame for the pollution degree of pathogenic Vibrio in marine products in China and established the dominant flora of pathogenic Vibrio in different types of aquatic products, which provides a theoretical basis for food safety supervision departments to take targeted prevention and control measures.

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Evaluation of loop-mediated isothermal amplification assay along with conventional and real-time PCR assay for sensitive detection of pathogenic Vibrio parahaemolyticus from seafood sample without enrichment

Cited by 14 publications

References 36 publications

Identification of the novel potential pathogen Trueperella pecoris with interspecies significance by LAMP diagnostics

Identification of the novel potential pathogen Trueperella pecoris with interspecies significance by LAMP diagnostics

Rapid, specific and sensitive detection of Vibrio parahaemolyticus in seafood by accelerated strand exchange amplification

Analysis of Pathogenic Vibrio Contamination in Marine Products along China Based on Fluorescence Quantitative PCR

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