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التنميط الجيني لفايروس التهاب الكبد نوع اي لمرضى الاصابة الحادة في محافظة المثنى - العراق == Genotyping of Hepatitis E Virus Among Patients With Acute Infection In Al - Muthanna Province / Iraq
Author name:
ازهار صبري مسلم الذهبي
Supervisor name:
رغد حربي مهدي العزاوي
General topic:
Biology
Specific topic:
Microbiology - Viruses
Degree:
Master
University:
University of Baghdad
Language:
English
University location:
Baghdad
First pages:
24T2998 - p.pdf
Abstract:
Hepatitis E Virus (HEV) infection is found worldwide. It is responsible for large outbreaks in East and South Asia. This study was carried out to identify the distribution of acute HEV infection in AlMuthanna province and then determine the genotypes responsible for the disease spreading. This is the first investigation about the occurrence of hepatitis E genotypes in Al - Muthanna province patients. The study included 270 jaundiced patients of both sexes From AlMuthanna province which included out patients in public health laboratory in Samawah city, for the period from October 2013 to June 2014. All these patients were tested for anti - HEV IgM antibodies by using enzyme linked immunosorbent assay (ELISA). In Al - Muthanna province, 72 patients (26.66 %) out of 270 patients showed positive results for anti - HEV IgM antibodies, and all those patients were tested for confirmatory test at central public health laboratories (CPHL) in Baghdad province. Those patients consist of 45 females and 27 males with age ranged between (4 - 74) years old, were all negative for routinely screened markers of Hepatitis A, Hepatitis B and Hepatitis C, Ten normal healthy individuals, used as normal control in this study. This study showed that the HEV IgM is more common among younger age group (15 - 24), with a percentage of) 41.67%(, and it was higher in females (63%) than in males (37%). Highly significant differences (p ? 0.01) appeared among age groups. The levels of liver function enzymes demonstrated significant differences (p ? 0.05) in HEV patients as compared with healthy controls. The levels of Total Serum Bilirubin (TSB) were (23.41 ± 12.38) mg/dl, Serum Aspartate transaminase (SAST) were (887.17 ± 9.87) U/l, Serum Alanine transaminase (SALT) were (859.91 ± 13.28) U/l, and the levels of Serum Alkaline phosphatase (SALP) were (206.56 ± 5.04) U/l higher than healthy controls (0.884 ± 0.05) mg/dl, (37.27 ± 4.58) U/l, (34.14 ± 7.63) U/l, (48.09 ± 15.42) U/l respectively. RNA was extracted from sera of positive anti - HEV IgM antibodies by using a QIAamp® Viral RNA Mini Kit. Nanodrop and Quantus™ Fluorometer instruments were used to determine the RNA yield and purity according to the standard kit protocol. High levels of RNA concentration were measured by Nanodrop and Quantus™ Fluorometer. Forty - six samples have high RNA concentration out of 72 samples were detected for genotypes I, III & IV by using Applied Biosystems Real time PCR 7500 machine. In this study, HEV type I and III were detected in 23 samples out of 46 samples by using Real Time PCR systems. Ten samples were positive for this test. Thirteen samples (56.52%) were negative for this test. The distribution percentage of genotypes I and III was (43.48%). The results appeared significant difference (P ? 0.05). Also by using the same technique (Real time PCR systems) the HEV type IV was detected in 23 samples out of 46 samples. Which included, 13 samples that recorded negative results by I & III genotypes kit and only 2 samples from it recorded positive results and 11 samples recorded negative result for HEV genotype IV. From other 10 samples, only 3 samples recorded positive result and 7 samples recorded negative result for HEV genotype IV. In a total, 5 samples were positive for HEV genotype IV and 18 samples (78.26%) were negative. So that the distribution percentage of genotype IV was (21.74%). In this study, the samples that showed negative results in both kits have not been tested for other genotypes. This study indicated that the acute HEV infection is highly endemic in Al - Muthanna province in Iraq. Genotypes I & III were the most distribution than genotype IV in patients from Al - Muthanna province in Iraq. These results suggest that the genotypes I & III are the main causative agents of sporadic HEV infection in Al - Muthanna province
