Dataset Information

Detection of Spiked Fasciola hepatica Eggs in Stool Specimens Using LAMP Technique.

ABSTRACT: Background:Fascioliasis is one of the most important food-borne worm disease caused by Fasciola sp. Parasitological diagnosis is more difficult due to the low parasite burden and a few eggs shedding of helminths. Therefore, it will be valuable to development of simple, fast and reliable diagnostic tests for detection of human and animal fascioliasis. Methods:Infected liver collected from abattoir in Tehran, Iran in 2017. F. hepatica eggs were detached from the uterus of worms under a stereo microscope. Various numbers of eggs were spiked to 200 mgr. of negative feces samples. DNA was extracted and then target regions (nuclear IGS) were amplified by LAMP assay using six primers. Fecal specimens without egg and DNA of other helminths were used as negative controls. F. hepatica sample which confirmed by morphologic criteria and PCR-RFLP was used as positive control. Results:LAMP products by using SYBR Green I could detect even a single egg in fecal samples which was visible by change of color from orange to green. There was no cross amplification by other helminths including; Taenia saginata, Dicrosolium dendriticum and F. gigantica. Conclusion:LAMP seems a rapid, sensitive, cost-effective technique for detection of human fascioliasis.

SUBMITTER: Ghodsian S

PROVIDER: S-EPMC6815858 | biostudies-literature | 2019 Jul-Sep

REPOSITORIES: biostudies-literature

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Detection of Spiked <i>Fasciola hepatica</i> Eggs in Stool Specimens Using LAMP Technique.

Ghodsian Sahar S Rouhani Soheila S Fallahi Shirzad S Seyyedtabaei Seyyed Javad SJ Taghipour Niloofar N

Iranian journal of parasitology 20190701 3

<h4>Background</h4>Fascioliasis is one of the most important food-borne worm disease caused by <i>Fasciola</i> sp. Parasitological diagnosis is more difficult due to the low parasite burden and a few eggs shedding of helminths. Therefore, it will be valuable to development of simple, fast and reliable diagnostic tests for detection of human and animal fascioliasis.<h4>Methods</h4>Infected liver collected from abattoir in Tehran, Iran in 2017. <i>F. hepatica</i> eggs were detached from the uterus ...[more]

PMID: 31673256

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Project description:Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop-mediated isothermal amplification (LAMP) technique with PCR assay for the diagnosis of F. hepatica in sheep. In this cross-sectional study, 195 stool samples were collected from sheep for 3 months in Lorestan province, West of Iran. Specimens' parasitological examination was performed by using the direct wet mount and formalin-ether concentration method. After DNA extraction from the samples, molecular analysis was done using PCR and LAMP techniques based on the Fasciola ribosomal intergenic spacer (IGS) sequence. Of 195 specimens of sheep, 11 specimens were identified as F. hepatica-positive infection by using microscopic, PCR and LAMP assays. Kappa agreement test results showed that there was a significant agreement between the results of microscopic examination diagnostic tests, PCR and LAMP (Kappa = 0.51-0.72 and p < .001). According to the results of chi-square comparisons between parasite prevalence applying different techniques and variables of age, sex breed, and type of drinking water, there was no significant relationship (p ≥ .05). However, most of the infected sheep with Fasciola were 3- to 4-year-old females, of the Lori breed and consumed tap water. In many endemic areas, successful prevention and treatment of fascioliasis in animals depend on rapid and accurate diagnosis. Based on the results of the Kappa agreement, the significant agreement among the results of the microscopic examination, PCR and LAMP indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in sheep. However, molecular methods, especially the LAMP technique, are suggested because of their higher sensitivity and reliability for the diagnosis of F. hepatica even under field conditions.

Dataset Information

Detection of Spiked Fasciola hepatica Eggs in Stool Specimens Using LAMP Technique.

Publications

Detection of Spiked <i>Fasciola hepatica</i> Eggs in Stool Specimens Using LAMP Technique.

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