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. 2018 Jun 10;6(6):982-987.
doi: 10.3889/oamjms.2018.227. eCollection 2018 Jun 20.

Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion

Amany A Abd El Aal  1   2 , Reham K Nahnoush  1 , Marwa A Elmallawany  1 , Walid S El-Sherbiny  3 , Mohamed S Badr  4 , Ghada M Nasr  5
Affiliations

Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion

Amany A Abd El Aal et al. Open Access Maced J Med Sci. .

Abstract

Aim: The current study aimed to assess the practicability of a simple loop-mediated isothermal amplification (LAMP) about real-time quantitative PCR to diagnose primary toxoplasmosis among high-risk pregnant women.

Methods: Cloned Toxoplasma samples were used to calculate the analytical sensitivity while specificity was assessed using pooled DNA samples extracted from other parasitic stages.

Results: Both techniques showed 100% sensitivity and specificity and then applied to detect recent Toxoplasma infection in peripheral blood of 77 IgG negative women out of a total 139 women lately experienced spontaneous abortion. The 2 techniques obtained positive results in 8 samples confirming primary toxoplasmosis.

Conclusion: Generally, LAMP assay is a simple, cost-effective molecular technique can be completed in less than half an hour to diagnose primary Toxoplasma infection. The technique can be applied in a minimally equipped laboratory by ordinary workers to screen the vulnerable groups. Further analysis using larger samples with the quantitative approach is recommended to confirm the sensitivity of this emergent molecular technique.

Keywords: Abortion; Isothermal; LAMP; Toxoplasma; qPCR.

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Figures

Figure 1
Figure 1
Samples in the upper photo show positive isothermal amplification for 8 samples (from 1 to 8) the sample number 9 is the negative control. The lower photo represents the real-time quantitative curves for the 8 positive samples with different crossing point at different cycles according to their genomic equivalent
See this image and copyright information in PMC

References

    1. Lopes FM, Gonçalves DD, Mitsuka-Breganó R, Freire RL, Navarr IT. Toxoplasma gondii infection in pregnancy. Braz J Infect Dis. 2007;11:496–506. https://doi.org/10.1590/S1413-86702007000500011 PMid:17962877. - PubMed
    1. Robbins JR, Zeldovich VB, Poukchanski A, Boothroyd JC, Bakardjiev AI. Tissue Barriers of the Human Placenta to Infection with Toxoplasma gondii. Infection and Immunity. 2012;80(1):418–428. https://doi.org/10.1128/IAI.05899-11 PMid:22083708 PMCid: PMC3255695. - PMC - PubMed
    1. Vado-Solís I, Suárez-Solís V, Jiménez-Delgadillo B, Zavala-Velázquez J, Segura- Correa J. Toxoplasma gondii Presence in Women with Spontanous Abortion in Yucatan, Mexico. 2013;99(2):383–385. - PubMed
    1. Sultana M, Sazzad Hossain Md, Dewan F, Sultan J, Rashid M. Association of Toxoplasma gondii Infection with Spontaneous Abortion. Bangladesh. J Obstet Gynaecol. 2014;29(2):87–93.
    1. Matin S, Shahbazi GH. Study on abortion associated with Toxoplasma gondii in women based on PCR detection of aborted placenta and maternal serology in Ardabil, Iran. J Med Microb Diagn. 2017;6:2.

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