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

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.

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

“…Both techniques were then employed to detect recent T. gondii infection in 77/139 IgG negative women who suffered from spontaneous abortion. The result showed that both techniques were positive in eight samples, thus confirming primary Toxoplasma infection [113].…”

“…The IAm Toxo was reported to be highly sensitive and robust and gave good results compared to the alternative amplification technologies [112]. The LAMP method has also been used to detect primary Toxoplasma infection in highrisk pregnant women with previous spontaneous abortion history [113]. The clinical sensitivity and specificity of LAMP were compared with quantitative real-time PCR.…”

“…Furthermore, no positive results were obtained in samples that were extracted from other parasites including Giardia, Cryptosporidium, Blastocystis, Leishmania spp., and Plasmodium spp. [113]. Both techniques were then employed to detect recent T. gondii infection in 77/139 IgG negative women who suffered from spontaneous abortion.…”

The global seroprevalence of anti-Toxoplasma gondii antibodies in women who had spontaneous abortion: A systematic review and meta-analysis

“…One study was not analyzed because it did not accurately quantify the number of IgM and IgG positive cases separately [48], and its data were only systematically presented; therefore, 71 papers were included in the meta-analysis. Eight articles were related to the seroprevalence of anti-T. gondii antibodies in women who had spontaneous abortion in present pregnancy [49][50][51][52][53][54][55][56]. Moreover, serologic methods were used in six of these studies [49-52, 55, 56].…”

Optimization of Sybr Green Quantitative Real Time Polymerase Chain Reaction (qPCR) using Excreted-Secreted Antigens (ESAs) Genetik Marker for Detection Toxoplasma gondii

“…Toxoplasma gondii is an obligate intracellular parasite, causing toxoplasmosis in almost all warm-blooded animals and humans worldwide. Toxoplasmosis is a zoonotic disease of serious public health concern (Opsteegh et al, 2015;El Aal et al, 2018). During T. gondii infection, the parasite releases a number of molecules called T. gondii excretory secretory antigen (ESAs) into the surrounding environment, which allows the organism to attack and survive inside the host cell through immunomodulation (Moncada and Montoya, 2012).…”