Rapid And Sensitive Detection Of <i>Lawsonia Intracellularis In Pigs</i> By Real-Time Loop-Mediated Isothermal Amplification

Park, Byung-Yong; Shim, Kwan Seob; Kim, Won-Il; Mukter, Hossain Md; Kim, Bumseok; Kwon, Jungkee; Park, Choi‐Kyu; Cho, Sung‐Jin; Jo, Inho; Cho, Ho-Seong

doi:10.1515/acve-2015-0002

Cited by 4 publications

(1 citation statement)

References 19 publications

(22 reference statements)

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“…Though the probe enhances the specificity of the RPA amplification, the probe is always hard to be found in a target gene and is expensive to be synthesized. Real-time loop-mediated isothermal amplification (RT-LAMP) has been developed for the detection of L. intracellularis [ 36 ]. However, the L. intracellularis RPA-LFD was faster than RT-LAMP (15 min in L. intracellularis RPA compared to 30–60 min in RT-LAMP).…”

Section: Discussionmentioning

confidence: 99%

Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick

Tian

Zhang

et al. 2019

BMC Vet Res

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BackgroundLawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim of this study was to develop a simple and rapid method for on-site detection of Lawsonia intracellularis (L. intracellularis). As the isothermal recombinase polymerase amplification (RPA) can be performed at a constant temperature and its product is directly observed on a lateral-flow dipstick (LFD) with naked eyes without electrophoresis, the RPA-LFD assay should be useful for field diagnosis of L. intracellularis as well as its detection from clinical samples.ResultsThe established RPA-LFD assay could be finished in 30 min at a wide temperature range of 25 to 40 °C, and the amplicons could be visualized by naked eyes. The developed RPA-LFD assay was specific to dnaA gene of L. intracellularis, and did not detect nucleic acids extracted from other common gastrointestinal pathogens. The minimum detection of this RPA-LFD method was 400 L. intracellularis per reaction, which was as sensitive as conventional PCR. Further, the RPA-LFD assay was performed with 150 clinical fecal samples and the detection results were compared with conventional PCR. Results showed that the coincidence rate of RPA-LFD and conventional PCR was 100%.ConclusionsThe combined RPA with LFD assay provides a simple, rapid, specific and sensitive alternative for field diagnosis of L. intracellularis infection.

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Section: Discussionmentioning

confidence: 99%

Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick

Tian

Zhang

et al. 2019

BMC Vet Res

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Haemorrhagic Bowel Syndrome in Fattenig Pigs

et al. 2016

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The aim of this work was to determine the possible causes of sudden death in fattening pigs associated with haemorrhagic bowel syndrome (HBS) in a commercial pig farm (Eastern Slovakia). The etiology and pathogenesis of haemorrhagic bowel syndrome is not clearly defi ned within the scientifi c community. For this reason, clinical examination and sampling from 90 kg fi nisher pigs was done. Samples of blood, faeces, and rectal swabs were taken for haematological, serological, bacteriological, virological, and parasitological examination. Necropsy and sampling was also conducted on the carcasses of 90 kg fi nisher pigs that died peracutely during the investigation period. Three weeks later, during slaughter of the same group of pigs, goal-directed investigation and sampling from the gastrointestinal system for histological examination was performed. Results show the presence of bloody diarrhoea caused by L. intracellularis, C. perfringens typ A and haemolytic E. coli. Infl ammation and loss of blood led to moderate normochromic (haemoglobin 12.85±0.95 g.dl-1) normocytic anaemia (erythrocytes 3.38±0.17 T.l-1), hydremia (haematocrit 0.17±0.01 l.l-1) and mild leukocytosis (18.7±4.68 G.l-1). Diagnostic necropsy confi rmed the picture of "haemorrhagic bowel syndrome" caused by infection and subsequent enterotoxemia without presence of gastric or intestinal displacement. Slaughter examination showed picture of proliferative ileitis typical for L. intracellularis. On the basis of our examinations, a major role in haemorrhagic bowel syndrome played infectious agents such as L. intracellularis, C. perfringens typ A and haemolytic E. coli. Negative fi ndings of salmonellosis, dysentery, feed mycotoxins, intestinal parasites, and pollution of drinking water exclude these factors from the etiology of haemorrhagic bowel syndrome.

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PREVALENCE OF ANTIBODY TO AND DNA OFLAWSONIA INTRACELLULARISIN SAMPLES FROM WILD ANIMALS IN KOREA

Hossain

Cho

2016

Journal of Wildlife Diseases

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We evaluated the prevalence of Lawsonia intracellularis infection in three wild animal species in Korea; the Korean water deer ( Hydropotes inermis ), Siberian roe deer ( Capreolus pygargus ), and raccoon dogs ( Nyctereutes procyonoides ). We collected 136 sera and 109 fecal samples from individuals in 10 Wildlife Rescue and Conservation Centers. Serum samples were tested for anti- L. intracellularis antibodies using a blocking enzyme-linked immunosorbent assay (bELISA), and fecal samples were subjected to a real-time PCR assay for L. intracellularis . Thirty-five (25.7%) sera and 36 (33.0%) fecal samples were positive. We found a higher proportion of positive sera (64.7%, χ=15.439, P<0.01) and feces (58.8%, χ=6.126, P<0.05) in raccoon dogs (χ=11.855, P<0.01) than in the other species (20% positive sera and 29% positive feces in Korean water deer; 20% positive sera and 25% positive feces in Siberian roe deer). Our data indicate infection by L. intracellularis in Korean water deer, Siberian roe deer, and raccoon dogs throughout the country. It is imperative to know whether these infected animal species are natural hosts for L. intracellularis in addition to domestic pigs ( Sus scrofa domesticus).

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Rapid And Sensitive Detection Of Lawsonia Intracellularis In Pigs By Real-Time Loop-Mediated Isothermal Amplification

Cited by 4 publications

References 19 publications

Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick

Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick

Haemorrhagic Bowel Syndrome in Fattenig Pigs

PREVALENCE OF ANTIBODY TO AND DNA OFLAWSONIA INTRACELLULARISIN SAMPLES FROM WILD ANIMALS IN KOREA

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