Molecular identification and phylogenetic analysis of Wuchereria bancrofti from human blood samples in Egypt.

Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.

GENETIC POLYMORPHISM OF TOXOPLASMA GONDII FROM RECENTLY INFECTED ABORTED EGYPTIAN WOMEN.

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.

MOSQUITO IDENTIFICATION AND MOLECULAR XENOMONITORING OF LYMPHATIC FILARIASIS IN SELECTED ENDEMIC AREAS IN GIZA AND QUALIOUBIYA GOVERNORATES, EGYPT.

Lymphatic filariasis is a vector-borne health problem that has been focally endemic in Egypt for centuries. The chief vectors of transmission are Culicinae species. Control measures in the form of mass drug administration of DEC citrate treatment have been implemented in Nile delta for almost a decade. This study aimed to identify the prevalent mosquito species in endemic areas in Giza and Qualioubiya governorates and to monitor Wuchereria bancrofti infection by detecting the parasite DNA in collected mosquitoes. Adult mosquitoes were collected using light traps hung indoors. Microscopic examination was performed to identify and examine the morphologic characters of mosquitoes. Female Culex mosquitoes were subjected to semi-nested PCR to detect filarial DNA targeting repetitive DNA sequences (pWbl2 repetitive region) specific for W. bancrofti. The results revealed 3 species of mosquitoes Culex pipiens, Culex pusillus and Culex quinquefasciatus with the predominance of Culex pipiens (85.7%). Wuchereria bancrofti DNA was not detected in any of the collected mosquito pools. With progress of elimination programme in Nile Delta, follow up studies with larger sample size are recommended as the predominance of Culex pipiens the main lymphatic filariasis vector remains a risk of transmission in such areas.