The comparison of polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) methods for detecting Fasciola hepatica in faecal samples of sheeps in Lorestan province

Subject Areas : Journal of Comparative Pathobiology

, S. Amiri ¹ , bahar shemshadi ² , , S Fallahi ³ , , S. Shirali ⁴

1 - .Department of Pathobiology, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 - Department of Pathobiology, Faculty of Veterinary Medicine, Azad University, Science and Research Branch, Tehran, Iran
3 - Department of Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran
4 - Department of Biotechnology, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran

Received: 2021-11-01 Accepted : 2021-11-01 Published : 2020-11-21

Keywords: Sensitivity, Diagnosis, digene, Parasite,

Abstract :

Fascioliasis is a zoonotic disease with long history in Iran. Different methods are used for detection of the disease and one of methods is LAMP. The present study was conducted to compare the LAMP and PCR methods for detection of Fasciola hepatica in fecal samples of sheeps in Lorestan province. During 3 months, 195 samples were collected by classical randomized method. The samples were stored in -20ºC for future investigations by LAMP and PCR. The samples were investigated by LAMP and PCR. The data were analyzed by SPSS software and Chi-square and Kappa coefficient was used. The results showed that LAMP technique showed significant difference with LAMP and detected infected samples (11 samples in LAMP and 7 samples in PCR). The results for kappa showed a strong agreement between the both methods with coefficient of 0.65 and P=0.001). The LAMP method showed higher sensitivity. In sum, LAMP method showed higher sensitivity compared to other methods and can be used for detection of F. hepatica.

References:

1-         Hosseini-Safa A. Rokni MB. Mosawi SH. Heydarian P. Azizi H. Davari A. et al. High-resolution melting analysis as an appropriate method to differentiate between Fasciola hepatica and F. gigantica. Iranian Journal of Public Health. 2019; 48: 501-507.

2-         John BC. Davies DR. Williams DJL. Hodgkinson JE. A review of our current understanding of parasite survival in silage and stored forages, with a focus on Fasciola hepatica metacercariae. Grass Forage Science. 2019; 74: 211–217.

3-         Chen MG. Mottm KE. Progress in assessment of morbidity due to Fasciola hepatica infection: a review of recent literature. Tropical Disease Bulletin. 1999; 87: 1-38.

4-         Zooghi E. Yousefivand J. Hajikhani R. Human fasioliasis in some countries. Ghalamestan Honar. 2004; P 12-15 [in persian].

5-         Lotfy WM. El-Morshedy HN. Abou El-Hoda, M. El-Tawila, M.M. Omar, 

E.A. & Farag, H.F. Identification of the Egyptian species of Fasciola. Veterinary Parasitology. 2002;103(4):323–32.Mercer DK, Stewart CS. Keratin hydrolysis by dermatophytes. Medical Mycology. 2019;57(1):13-22.

6-         Martínez-Valladares M. Rojo-Vázquez FA. Loop-mediated isothermal amplification (LAMP) assay for the diagnosis of fasciolosis in sheep and its application under field conditions. Parasites & Vectors. 2016; 9:73.

7-         Notomi T. Okayama H. Masubuchi H. Yonekawa T. Watanabe, K. Amino, N. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research. 2000; 28(12): E63.

8-         Krasteva D. Toubiana M. Hartati S. Development of loop-mediated isothermal amplification (LAMP) as a diagnostic tool of toxoplasmosis. Veterinary Parasitology. 2009; 162: 327–331.

9-         Kialashaki E. Sharif M. Fakhar M. Daryani A. Paghe AS. Assessing Different Methods of DNA Extraction from Giardia Lamblia Cysts. Journal Mazandaran University Medical Sciences. 2013; 23(98): 67-74. [in persian].

10-       Ai L. Li C. Elsheikha HM. Hong SJ. Chen JX. Chen SH. et al. Rapid identification and differentiation of Fasciola hepatica and Fasciola gigantica by a loop-mediated isothermal amplification (LAMP) assay. Veterinary Parasitology. 2010; 174(3-4): 228-233.

11-       Mizani A. Gil P. Sarvari Sh. Daryani A. Sharif M. Amooei A. et al. Detection and differentiation of Fasiola hepatica and Fasiola gigantica by loop-mediated Isothermal Amplification. Mazandaran Medical University Journal. 2016; 145:45-53. [in persian].

12-       Mizani A. Gill P. Sarvi Sh. Daryani A. Sharif M. Rahimi MT. et al.  Journal Mazandaran University Medical Sciences. 2015; 25(122): 72-80. [in persian].

13-       Dalton JP. Robinson MW. Mulcahy G. O’Neill SM. Donnelly S. Immunomodulatory molecules of Fasciola hepatica: Candidates for both vaccine and immunotherapeutic development. Veterinary Parasitology. 2013; 195: 272–285. https ://doi.org/10.1016/j. vetpar.2013.04.008.

14-       Ni XW. McManus DP. Lou ZZ. Yang JF. Yan HB. Li L. et al. A comparison of Loop-Mediated Isothermal Amplification (LAMP) with other surveillance tools for Echinococcus granulosus diagnosis in canine definitive hosts. Parasites & Vectors. 2014; 9(7): 254.

15-       Kazemi F. Arjmand R. Tavalla M. Prevalence of toxocara species in park soils of Ahvaz City, southwest of Iran. Asian Pacific Journal of Tropical Disease. 2017; 7(12): 705-707. https://doi.org/10.12980/apjtd.7.2017D7-173

16-       Pazooki Ghoohe H. Haghparast B. Fakhar M. Common and novel methods for laboratory detection and identification of Leishmaniosis. Mazandaran Medical University Journal. 2015; 132; 345-361.

17-       Kudyba HM. Louzada J. Ljolje D. Kudyba K. Muralidharan V. Oliveira‑Ferreira J. et al. Field evaluation of malaria malachite green loop‑mediated isothermal amplification in health posts in Roraima state, Brazil. Malaria Journal. 2019; 18: 98-105. https://doi.org/10.1186/s12936-019-2722-1.

18-       Ezatpour B. Hasanvand A. Azami M.  Anbari Kh. Ahmadpour F. Prevalence of liver fluke infections in slaughtered animals in Lorestan Iran. Journal  of Parasitic Disease. 2015; 39, 725–729. https://doi.org/10.1007/s12639-014-0428-4