Badr Mohamed S, Attia Samar S, El-Sherbiny Walid S, Abd-Allah M A El Ebidi, Hefny Hesham M, Salem Ahmed N M
J Egypt Soc Parasitol. 2016 Apr;46(1):49-55. doi: 10.12816/0026149.
Toxoplasmosis caused by Toxoplasma gondii is one of the most prevalent parasitic diseases in human beings. Human toxoplasmosis can be associated with serious clinical manifestations, particularly in developing fetus. The aim of the current study was to identify the possible lineage type of Toxoplasma gondii, molecularly detected in placental samples of women whose pregnancies were spontaneously terminated in the first trimester. Preliminary detection of Toxoplasma genomic materials was done by a SYBR green qPCR technology. Subsequent identification of Toxoplasma strain was done for the positive samples using PCR-restriction fragment length polymorphism (RFLP) at the SAG2 loci of T. gondii using restriction enzymes HhaI and Sau3AI. Out of 72 tested samples, Toxoplasma B1 gene was detected in 9 cases. Toxoplasma genotypes I and II in addition to unknown type were identified in 4, 3 and 2 cases respectively, while type III was not detected in our samples, hence excluded as a leading cause of abortion in humans in our preliminary study. Nevertheless, it remains uncertain to what extent the genotype of the parasite directly contributes to the clinical severity of human toxoplasmosis. Certainly, advanced molecular techniques targeting different Toxoplasma strains are crucial for better understanding of human toxoplasmosis. For more elucidation, additional studies are recommended intended for genetic characterization of such serious parasitic infection using larger number of samples.
由刚地弓形虫引起的弓形虫病是人类最常见的寄生虫病之一。人类弓形虫病可能伴有严重的临床表现,尤其是在发育中的胎儿身上。本研究的目的是鉴定在孕早期自然流产女性的胎盘样本中分子检测到的刚地弓形虫可能的谱系类型。通过SYBR Green qPCR技术对弓形虫基因组材料进行初步检测。使用限制性内切酶HhaI和Sau3AI,通过PCR-限制性片段长度多态性(RFLP)在刚地弓形虫的SAG2位点对阳性样本进行后续的弓形虫菌株鉴定。在72个检测样本中,9个样本检测到弓形虫B1基因。分别在4例、3例和2例样本中鉴定出弓形虫基因型I、II以及未知类型,而在我们的样本中未检测到III型,因此在我们的初步研究中排除其作为人类流产主要原因的可能性。然而,尚不确定寄生虫的基因型在多大程度上直接导致人类弓形虫病的临床严重程度。当然,针对不同弓形虫菌株的先进分子技术对于更好地理解人类弓形虫病至关重要。为了进一步阐明,建议使用更多样本对这种严重寄生虫感染进行基因特征分析的后续研究。