Imported malaria in United Arab Emirates: evaluation of a new DNA extraction technique using nested PCR.

Local malaria transmission in the United Arab Emirates (UAE) came to an end in 1997. Nevertheless, UAE has been subjected to substantial importation of malaria cases from abroad, concerning both UAE nationals and immigrants from malarious countries with a total number of 2,119 cases in 2007. To evaluate a new DNA extraction technique using nested PCR, blood samples were collected from 132 individuals who presented to Infectious Diseases Department in Rashid Hospital, Dubai, and Central Department of Malaria Control with fever and persistent headache. Giemsa-stained blood films and ELISA test for malaria antibodies were carried out for detection of Plasmodium infection. Plasmodium infections were identified with the genus-specific primer set and species differentiation using nested PCR. A rapid procedure for diagnosis of malaria infections directly from dried blood spots using for the first time DNA extract from FTA Elute cards was evaluated in contrast to extraction techniques using FTA classic cards and rapid boiling technique. Our new simple technique for DNA extraction using FTA Elute cards was very sensitive giving a sensitivity of 100% compared to 94% using FTA classic cards and 62% in the rapid boiling technique. No complex preparation of blood samples was required prior to the amplification. The production cost of DNA isolation in our PCR assay was much less in comparable to that of other DNA extraction protocols. The nested PCR detected plasmodial infection and could differentiate P. falciparum from P. vivax, and also detected the mixed infection.

Genetic characterization of Giardia lamblia isolates from Egyptian patients with relation to clinical giardiasis.

Investigators tried to correlate clinical presentation of giardiasis to the different genotypes, but controversial data were described through the last decade. The clinical manifestations of 89 Giardia patients were classified into:- GI: 52 symptomatic patients and GII: 37 asymptomatic patients. Genetic characterization of G. lamblia of the patients' fecal samples was performed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) technique b using triose phosphate isomerase gene (tpi). Forty-two patients had genotype A1 (47.19%), 16 had genotype A2 (17.98%), 10 had genotype B (11.23%) and 19 had mixed genotype infection (21.35%). However, PCR-RFLP failed to determine Giardia genotype of only two cases (2.25%). The 20 control fecal samples obtained from healthy subjects showed negative results for G. lamblia by PCR-RFLP method. Of 52 cases in GI, the prevalence genotype A1 was 44.23%, genotype A2 was 19.23%, genotype B was 13.46%, mixed genotype infection was 21.15% and undetermined genotype was 1.92% as compared to 51.35%, 16.21%, 8.11%, 21.62% and 2.70% in GII, respecttively. There was no significant difference between both groups as regard the different Giardia genotypes (p>0.05). Statistical analysis of each symptom in different genotypes revealed insignificant (p>0.05). The results denied any correlation between G. lamblia genotype and the clinical presentation of giardiasis.

Intestinal parasites in Dakahlia governorate, with different techniques in diagnosing protozoa.

A total of 3180 patients attending Mansoura University Hospitals' Clinics, were subjected to stool examination by direct wet smear, formol-ether concentration, original formol-tween concentration, modified formol-tween concentration, modified Sheather's sugar floatation, Potassium hydroxide concentration and Gomori's Trichrome stain, and modified Kinyoun's acid-fast stain, and Ryan's Trichrome blue stain for Microsporidia. The intestinal helminthes in a descending order of abundance were: S. mansoni (5.3%), Fasciola sp. (4.8%), H. heterophyes (4.2%), Hymenolepis nana (3.9%), Trichostrongylus sp. (2.6%), A. lumbricoides (1.8%), Strongyloides stercoralis (1.5%), H. diminuta (1.4%), Taenia saginata (1.1%), E. vermicularis (by smear; 1.1 %), T. trichura (0.7%) and lastly A. duodenale (0.1%). The intestinal protozoa in a descending order of abundance were Blastocystis hominis (22.4%), Giardia lamblia (19.6%), Entamoeba histolytica/E.dispar (19%), Iodamoeba butschlii (16%), Cryptosporidium parvum (14.3%), E. coli (9.7%), Isospora hominis (7.7%), Endolimax nana (6.9%), E. hartmani (5.9%), Dientamoeba fragilis (5.1%), Chilomastix mesnili (5.1%), Trichomonas hominis (4.2%), Cyclospora cayetanensis (4.2%), Microsporidia spores (3.2%), Enteromonas hominis (1.9%) and Embadomonas intestinalis (1.3%). The results were discussed.

Hydatidosis granulosus in Egyptian slaughtered animals in the years 2000-2005.

Hydatidosis was investigated among camels, sheep, goats, and pigs in Egyptian official abattoirs, from August 2000 to August 2005, and among cows and buffaloes were in Mansoura official abattoirs, in the year 2005. One hundred randomly chosen animals of each species were subjected to serologic and histopathologic examinations for infections. The overall five years hydatidosis prevalence was 2.53%, 0.3% & 0.68% in camels, sheep & goats, and pigs respectively. The 2005 year prevalence in cows and buffaloes was 6.4% in Mansoura. There was a significant difference between animals regarding liver infection, but the difference was highly significant for lung infection. A highly significant difference in hydatid cysts size was between pigs and other animals species (p< 0.000) and a significant difference between macroscopic findings in pigs and camels (p=0.018). A high significant difference was between histopathology in all animals species except pigs and sheep & goats (p=0.089). IHAT showed highly significant difference between camels and other animals species (p<0.000). A significant histopathologic positive correlation was between positive IHAT and fertility (Pearson correlation =0.148, p=0.003). The results were photographed and discussed.

The reflection of control programs of parasitic diseases upon gastrointestinal helminthiasis in Dakahlia Governorate, Egypt.

The study area included Mansoura city as an urban area and Gogar village as a rural area. One thousand individuals were randomly selected from each area. Different methods of stool examination, perianal swab and urine examination of all participants revealed that the incidence in Mansoura city was in a descending order Heterophyes heterophyes 6.4%; Enterobius vermicularis 3.9%; Hymenienolepis nana 2.2%; Schistosoma mansoni 0.5%; Trichostrongylus colubriftormis; Strongyloides stercoralis and Fasciola sp. were recorded as 0.2% of each. Taenia saginata, Ascaris lumbricoides and Trichocephalus trichiuris were recorded as 0.1% of each. Neither Ancylostoma duodenale nor Hymenolepis dimninuta was recorded. In Gogar, the parasitic infection was H. hetephyes 4.5%; E. vermicularis 4.1%: H. nana 3.3%; S. mansoni 1.6%; T. colubriformis 0.9%; S. stercoralis 0.5%. Fasciola sp. 0.4%; T. saginata, A. lumbricoides, H. diminuta, A. duodenale and T. trichiuris were recorded as 0.1% of each. None S. haematobiumn was detected in both areas. So, the infection rates of H. heterophyes, E. vermicularis, H. nana S. mansoni, Fasciola sp., T. colubriformis and S. stercoralis were relatively high the rural than in urban area. This was not surprising since the socioeconomic, hygienic conditions and medical services were relative high in the city than in the village. No doubt, the identifications of parasitosis pave the way for feasible treatment and control measures.

Evaluation of seven assays detecting serum immunoglobulin classes and subclasses and salivary and faecal secretory IgA against Fasciola excretory/secretory (ES) antigens in diagnosing fascioliasis.

Seven assays detecting serum IgM, IgG, IgG1, IgG4, IgA and salivary and fecal excretory IgA against Fasciola excretory/secretory (ES) antigens were evaluated in diagnosing fascioliasis, for cross reactivity with Schistosoma mansoni sera and for evaluation of cure of Fasciola infection after treatment. Assays detecting sera IgM, IgG1, IgG4 and IgA against Fasciola ES antigens showed 100% specificity and sensitivity. Assays detecting IgM and IgG showed 98% and 96% sensitivity and 100% and 94.6% specificity respectively. Assays detecting salivary and faecal IgA showed 92% & 96% sensitivity and 100% & 100% specificity respectively. Assays detecting IgM and IgG4 were the best in evaluation of cure and assays detecting IgG4 & IgA showed the lowest cross-reactivity with sera from S. mansoni infected patients. So, assays detecting serum IgA, IgG1 & IgG4 against Fasciola ES antigens were highly sensitive and specific for diagnosis of fascioliasis and assays detecting salivary and faecal IgA were promising and of great help in diagnosis of fascioliasis especially in epidemiologic studies.