Feasibility of a DNA biosensor assay based on loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the visual detection of Ascaridia galli eggs in faecal samples.
{"title":"Feasibility of a DNA biosensor assay based on loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the visual detection of <i>Ascaridia galli</i> eggs in faecal samples.","authors":"Wasin Panich, Thanawan Tejangkura, Thapana Chontananarth","doi":"10.1080/03079457.2023.2196251","DOIUrl":null,"url":null,"abstract":"<p><p><i>Ascaridia galli</i> is an important nematode that causes ascaridiasis in free-range and indoor system chicken farms. Infection with <i>A. galli</i> may damage the intestinal mucosa and inhibit nutrient absorption, leading to a reduced growth rate, weight loss and a decreased egg production. Consequently, <i>A. galli</i> infection is a significant health problem in chickens. In this study, we developed a loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay for the visual detection of <i>A. galli</i> eggs in faecal samples. The LAMP-LFD assay consists of six primers and one DNA probe that recognize the internal transcribed spacer 2 (ITS2) region; it can be performed within 70 min and the results can be interpreted with the naked eye. Using the LAMP-LFD assay developed in this study, <i>A. galli</i> DNA was specifically amplified without any cross-reactions with other related parasites (<i>Heterakis gallinarum</i>, <i>Raillietina echinobothrida</i>, <i>R. tetragona</i>, <i>R. cesticillus</i>, <i>Cotugnia</i> sp., <i>Echinostoma miyagawai</i>) and definitive hosts (<i>Gallus gallus domesticus</i>, <i>Anas platyrhynchos domesticus</i>). The minimum detectable DNA concentration was 5 pg/μl, and the detectable egg count was 50 eggs per reaction. The assay can be performed in a water bath, without the need for <i>post-mortem</i> morphological investigations and laboratory instruments. It is therefore a viable alternative for the detection of <i>A. galli</i> in chicken faeces and can replace classical methods in field screening for epidemiological investigations, veterinary health and poultry farming management.<b>RESEARCH HIGHLIGHTS</b>This is the first study using the LAMP-LFD assay for <i>Ascaridia galli</i> detection.The results can be observed by the naked eye.The developed assay can be used to detect <i>Ascaridia galli</i> eggs in faecal samples.</p>","PeriodicalId":8788,"journal":{"name":"Avian Pathology","volume":null,"pages":null},"PeriodicalIF":2.5000,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Avian Pathology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1080/03079457.2023.2196251","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"VETERINARY SCIENCES","Score":null,"Total":0}
引用次数: 2
Abstract
Ascaridia galli is an important nematode that causes ascaridiasis in free-range and indoor system chicken farms. Infection with A. galli may damage the intestinal mucosa and inhibit nutrient absorption, leading to a reduced growth rate, weight loss and a decreased egg production. Consequently, A. galli infection is a significant health problem in chickens. In this study, we developed a loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay for the visual detection of A. galli eggs in faecal samples. The LAMP-LFD assay consists of six primers and one DNA probe that recognize the internal transcribed spacer 2 (ITS2) region; it can be performed within 70 min and the results can be interpreted with the naked eye. Using the LAMP-LFD assay developed in this study, A. galli DNA was specifically amplified without any cross-reactions with other related parasites (Heterakis gallinarum, Raillietina echinobothrida, R. tetragona, R. cesticillus, Cotugnia sp., Echinostoma miyagawai) and definitive hosts (Gallus gallus domesticus, Anas platyrhynchos domesticus). The minimum detectable DNA concentration was 5 pg/μl, and the detectable egg count was 50 eggs per reaction. The assay can be performed in a water bath, without the need for post-mortem morphological investigations and laboratory instruments. It is therefore a viable alternative for the detection of A. galli in chicken faeces and can replace classical methods in field screening for epidemiological investigations, veterinary health and poultry farming management.RESEARCH HIGHLIGHTSThis is the first study using the LAMP-LFD assay for Ascaridia galli detection.The results can be observed by the naked eye.The developed assay can be used to detect Ascaridia galli eggs in faecal samples.
期刊介绍:
Avian Pathology is the official journal of the World Veterinary Poultry Association and, since its first publication in 1972, has been a leading international journal for poultry disease scientists. It publishes material relevant to the entire field of infectious and non-infectious diseases of poultry and other birds. Accepted manuscripts will contribute novel data of interest to an international readership and will add significantly to knowledge and understanding of diseases, old or new. Subject areas include pathology, diagnosis, detection and characterisation of pathogens, infections of possible zoonotic importance, epidemiology, innate and immune responses, vaccines, gene sequences, genetics in relation to disease and physiological and biochemical changes in response to disease. First and subsequent reports of well-recognized diseases within a country are not acceptable unless they also include substantial new information about the disease or pathogen. Manuscripts on wild or pet birds should describe disease or pathogens in a significant number of birds, recognizing/suggesting serious potential impact on that species or that the disease or pathogen is of demonstrable relevance to poultry. Manuscripts on food-borne microorganisms acquired during or after processing, and those that catalogue the occurrence or properties of microorganisms, are unlikely to be considered for publication in the absence of data linking them to avian disease.