Rapid diagnosis of Toxoplasma gondii using loop-mediated isothermal amplification assay in camels and small ruminants

Gamil S G Zeedan,Amira H El-Namaky,Abeer M Abdalhamed,Raafat M Shaapan

doi:10.1186/s43088-021-00184-x

Gamil S G Zeedan, Amira H El-Namaky + Show 2 more

Open Access

https://doi.org/10.1186/s43088-021-00184-x

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Abstract

BackgroundThis study was conducted to detect the presence of T. gondii in milk and blood samples using three different assays: enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification assay (LAMP). Whole blood, serum, and milk samples were collected from goats (n = 156), sheep (n = 261), and camels (n = 108) in different governorates in Egypt from December 2019 to February 2021 and screened by ELISA for anti-Toxoplasma IgG antibodies before DNA extraction. The target T. gondii DNA gene was detected and evaluated using the LAMP assay compared to PCR.ResultsT. gondii antibodies were found in milk and serum samples at the rates of (29.26%) and (36.58%) in camels, (34.18%) and (35.89%) in sheep, and (33.7%) and (36.36%) in goats, respectively. Similar to PCR, the percentages of LAMP tests for the detection of the T. gondii DNA gene in milk and blood samples of camels, sheep, and goats were (4.8, 14.63), (6.83, 7.69), and (7.79, 9.09), respectively. LAMP's sensitivity for detecting T. gondii in milk and blood samples, which was identical to that of PCR, was 100%.ConclusionsThe findings clearly demonstrated that there were no variations in T. gondii detection capabilities in milk and blood samples from various animals using both PCR and LAMP tests. It provides a quick, precise, and sensitive method of detecting T. gondii in a variety of samples that may be used both in the field and in laboratory diagnosis.

Highlights

This study was conducted to detect the presence of T. gondii in milk and blood samples using three different assays: enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification assay (LAMP)
The prevalence of T. gondii was higher in male goats than in sheep; 18/31 (58%) and 15/31 (48.3%), respectively
The present study revealed that the high incidence of T. gondii in lactating goats’ and lactating sheep’s milk, but not in lactating camel milk, is due to the difference between the types of pastures of small ruminates and the camels in different governorates as shown in Figs. 1, 2, and 3

Summary

Introduction

This study was conducted to detect the presence of T. gondii in milk and blood samples using three different assays: enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification assay (LAMP). Serum, and milk samples were collected from goats (n = 156), sheep (n = 261), and camels (n = 108) in different governorates in Egypt from December 2019 to February 2021 and screened by ELISA for anti-Toxoplasma IgG antibodies before DNA extraction. The real-time PCR (qPCR) technique is well known for its concurrent detection and quantification of extracted T. gondii–DNA gene amplicon. This technique has significantly shortened time and has comparable performance to cPCR, which is machine-dependent, often expensive, and requires regular maintenance

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