Author name:
مها اسماعيل مصطفى الجبوري
Supervisor name:
يحيى جرجيس سلمان
Abstract:
The current study had been carried out from January 2013 to July 2013 in medical laboratory researches - Kirkuk Faculty of Medicine. A total of 310 stool samples have been collected from children suffering from liquid diarrhea, their ages are from below one year to 12 years; six different laboratory diagnostic methods have been applied for detecting Giardia lamblia and other intestinal parasites. For microscopy diagnosis; direct wet double prepartions, zinc sulphate flotation are used. While immunological methods involve; Enzyme Linked Immuno - Sorbent Assay, corpo - antigen (ELISA), Direct Fluorescent Assay (DFA) and Lateral immune - chromatography assay (Triage panel). Giardia genome amplification is done using conventional Polymerase Chain Reaction (PCR) single step procedure by using of mixed primers of Giardia assemblages A1, A2 and B. The total rate of intestinal parasitic infection is 51.93 % distributed in 161 stool samples. This rate involve high frequency of protozoan infection in 132 (42.58 %) compare to 29 (9.35 %) for helminthes, P<0.05. More common intestinal protozoan parasites were Giardia lamblia 63(20.32 %) followed by Blastocyst hominis, Cryptosporidium parvum, Entamoeba coli, Entamoeba histolytica, Iodomoeba butschili, Endolimax nana and Balantidium coli with the rates : 6.77%, 6.45%, 4.18 %, 2.58 %, 1.29 %, 0.64 % and 0.32 % receptively. Concerning intestinal helminthic infection, high number and rate 23(7.41 %) was with Hymenolepis nana is compare with 1(0.32 %) record for Ancylostoma duodenale.According to gender high rate of giardiasis is recorded among males than in females, conversely to high rate of other intestinal parasites among female than in males P<0.05. Statistically relationship between Giardia distribution and ages are not significant P>0.05. Giardia co - existence are highly detected with Balstocystis hominis, and Cryptosporidium parvum total Giardia mixed parasitic infection rate 8.06 % is lower than12.23 %of pure Giardia lamblia infection, P>0.05. Regarding Giardia lamblia detecting according to laboratory methods; high rate of giardiasis 22.29% is reported by using PCR technique, followed by 20, 23 % by using direct wet preparation technique, P<0.05.The efficacy of laboratory methods for detecting Giardia stages are reduced; the following rates 19.35%, 19.03%, 17.74% and 14.51% obtained by using ELISA, DFA, flotationand lateral immuno - chromatography assay (Triage) respectively, P<0.05. Also statistical analysis reveals significance of PCR sensitivity, specificity, and accuracy in detecting giardiasis than other laboratory methods. Negative predictive values NPV in relation to type of laboratory methods are high, but statistically they are not significant, controversy to positive predictive value PPV that are significant. The efficacy of Triage panel is high for detecting giardiasis 14.45 % as compare with 4.5 % and 3.5 % for detecting Cryptosporidium parvum oocysts and Entamoeba histolytica respectively P<0.05 %. Considering the application of double direct wet preparations, the results of using this method are beneficial for detecting protozoan and helminthic parasites. While the using of zinc sulphate flotation technique reveal fluctuated results in spite of significant statistical analysis P<0.05. The employee of five laboratory methods for detecting the oocysts of Cryptosporidum parvum; the following rates 6.45%, 5.48%, 4.83%, 4.51% and 3.22% are recorded with the usage of DFA, direct microscopy, modified Ziehl - Neelsen method, Traige panel and flotation method respectively, P>0.05. Giardia lamblia DNA extraction from 80 stool samples that amplified using Giardia gene loci K725, reveal 66 samples positivity, pure Giardia lamblia genomic mass mean is 437.56 bps, with 1.705 % of genome purity and the extension of genomes range from 280 to750bps.While 23 of mixed Giardia plus other protozoan parasites, the mean of gemonic mass is 439.89 bps with genome purity mean 1.56 %. Amplification of Giardia genome mixed with helminthes reveal 443.33 bps of genomic mass mean and 1.49 % genome purity mean. In general the all genomic mass of Giardia lamblia (Pure and mixed infection) is 440.26 bps and purity mean 1.54 % P>0.05.PCR amplification in stool sample exert that Giardia lamblia genomes are mixed of human and animal type.
