Incidence of hepatitis A virus among hospitalized children's in Cairo

Hepatitis A virus [HAV] is an important cause of acute hepatitis worldwide that can lead to severe illness or even death. It is transmitted by the fecal-oral route through the consumption of contaminated food or water. This study was carried out to determine the incidence of HAV infection among hospitalized children's with acute hepatitis and genotyping of HAV Strains circulating in Greater Cairo. To fulfill the aim of the work, stool samples were collected from 102 hospitalized Children's, ages ranged from 0.5 to 12 years during the period from Dec. 2007 to Nov. 2008. Collected stool samples were submitted to nested RT-PCR for amplification of the VP1/2A region of the HAV genome. The expected fragment sizes of PCR products were 391bp and 244 bp for the first and second round of PCR, respectively. PCR products, of 2nd round of some positive samples, were purified for nucleotide sequence analysis in both directions. Fragments nucleotide sequences were compared to sequences derived from the corresponding HAV genome regions in the Gen Bank. Obtained data showed that HAV RNA prevalence were 82.35% [84/102] among hospitalized children's with acute hepatitis, and the highest HAV RNA was in the age group 3-5 and 9-12, where it reached 88.88% [40/45] and 81.8% [18/22] respectively, and the lowest prevalence rate was 70% [7/10] in the age group 0.5-2. Also, the incidence of HAV RNAwas higher in females 88.1%[37/42], than in males 78.33% [47/60]. Seasonal variation of HAV revealed that the viral incidence was 100% during winter and spring seasons, [25/25] and [17/17], respectively. While it was 85.36% [35/41] and 36.84% [7/19] in summer and autumn, respectively. Sequence analysis of selected fragments showed that all fragments are the same isolate. The phylogenic tree of positive samples confirmed that the isolated virus sequencing was most closely related to Hepatitis A virus isolate Egypt-swS5 deposited in the Gen Bank by accession no. [FJ0100837.2], with 100% of nucleotide identity

Epidemiology of rotavirus in greater Cairo

Rotaviruses are a major cause of gastroenteritis in children worldwide. This study was carried out to determine the incidence of rotavirus infection among children and the genotypes circulating in Greater Cairo. During May 2006 to April 2008 one hundred and sixty eight wastewater samples and five hundreds and sixty seven stool specimens from children with acute diarrhea were examined using RT-PCR and multiplex nested PCR for genotyping. Prevalence of rotavirus reached 162/567 [28.6%] and 10/48 [20.8%] in fecal specimens and raw wastewater samples, respectively. A marked occurrence of rotaviruses appeared during autumn and winter [from Sept. to Feb.]. In stool samples, the G-P type combination were G1P[4] 14.8%, G1P[6] 9.9%, G3P[4/ ] 9.9% and G3P[6] 8, 6%, while in wastewater samples G1P[4] 20.6%, and G3P[4] 5.9% were only detected. The efficiency of Zenin wastewater treatment plant [WWTP] to remove infectious rotavirus was higher than El Gabal El Asfar [WWTP] as the former uses a final chlorination step. The time needed for complete removal of the rotavirus infectious units and its genome was 15 and 30 mm, respectively when exposed to UV Lamp[Vilber-Lour mat T-15c] 9W and a virus concentration of 2x 106 CC-RT-PCR units/ml, The chlorine dose 15 mg for 30 mm is sufficient to remove the infectious units of rotavirus at 2x106 CC-RT-PCR units /ml, while the same dose for 45 mm is needed for complete removal of the rotavirus genome. The antiviral activity of a pigenin 7-o-glucoside [extracted from Chrysanthemum coronarium] against rotavirus was 60%

Molecular characterization of rotavirus infection among Egyptian children

Rotavirus is the major etiologic agent of severe diarrhea in children world wide. To study the incidence of rotavirus genotypes circulating in Egypt, one hundred fecal specimens were collected from hospitalized children's with acute diarrhea during Feb.2009 to Jan.2010. Using semi nested multiplex RT-PCR for both 0 and P typing. Rotavirus prevalence was 19% [19/100]. Seasonal distribution of rotavirus was 32% [8/25], 28% [7/25], 12% [3/25], and 4% [1/25] in winter, summer, autumn and spring, respectively. G I-genotype was the most predominant and frequent in all samples, followed by G4 and [19. This study revealed that P-genotypes were rare [5/100]. The detected P-genotypes were restricted to p[l I], p[4], and p[8J. A total of 881bp PCR fragments were sequenced and compared to sequences derived from the corresponding rotavirus genome deposited in GenBank. Phylogenetic tree of Rotavirus segment 9 isolated confirmed that the isolated virus sequencing was most closely related to the previously published sequences from different localities all over the world especially that isolated from Russia, South Africa and, Italy with 97-99% homology. A marked seasonal occurrence of rotavirus, during summer and winter season. G1-genotype was the most predominant and frequent in all samples while P-genotypes were restricted to P[l1], P[8]and P[4]

Coxsackie B Viruses as the main etiologic agent of myocarditis: Characterization of capsid proteins

Enteroviruses especially Coxsackie B viruses [CBVs] are responsible for approximately 50% of viral myocarditis cases. Coxsackie B viruses were inoculated in BGM cells for three successive passages. Viral capsid proteins were screened by SDS-PAGE, followed by Western blot. Electrophoretically separated proteins of CB2, CB4 in comparison to CA9, Echo9, and PV1 Sabin, showed common peptides particularly at the low molecular weight [ranging between 40 to 10 kDa]. The results of immunoblotting analysis using polyclonal antibodies raised against the whole Coxsackieviruses B viral particles did not reflect that all those bands are immunogenic, nevertheless those peptides could be used to raise specific antibodies which in turn could be useful in detecting viral circulating antigens

TT Virus outbreak among workers in wastewater treatment plants

The role of TT virus [TTV] as a human pathogen and the mode of transmission are unclear. To determine the prevalence of TTV infection and possible fecal-oral route of transmission, 24 wastewater samples from wastewater treatment plants was collected. Fecal samples were obtained from 60 workers in twaste water treatment plants and 15 healthy persons; all fecal samples were from persons non-transfused blood or blood products. TTV in tested samples were detected by nested PCR, using the NG primer sets. DNA sequences were analyzed from PCR products in both directions. TTV-DNA in water samples was 12.5% [3/24]. While, the overall prevalence of TTV in fecal excretion were 11.66% [7/60], and 6.6% [1/15] in workers and control group; respectively. A total of 100 bp PCR fragments were sequenced and compared to sequences derived from the corresponding TTV genome region deposited in GenBank. Present sequencing was most closely related to TTV-like mini virus, complete genome [Accession NC 002195], at nucleotide number 955 to 1021, suggesting related environmental sources of TTV infection. The phylogenetic analysis suggested that present strain might be sub-strain or mutation from the parent gene of TTV-like mini virus. The present of TTV in wastewater workers may be due to the interaction with contaminated environment and increased susceptibility to infectious agents

Waterborne viruses associated with repeated abortion

In this study we tried to find the role of some waterborne viruses in repeated abortion of women. The study includes maternal blood serum and fetal tissue. The serum of full term delivered women was taken as a control. All collected samples were inoculated on BGM and Hep2G cells to detect entero and Hepatitis E viruses. Enzyme-linked immunosorbent assay was also carried out for IgM and IgG antibodies against HEV in all serum samples. HEV-Ag was determined by dot-ELISA, which used also for enterovirus typing. Reverse transcriptase polymerase chain reaction was used for detection of entero and HE virus RNAs in the collected serum samples. To follow up the source of virus transmission, the wastewater treatment plant which serves the area of samples population was studied at the intake and the final effluent for the presence of hepatitis E virus and enteroviruses with special reference to coxsackieviruses. Wastewater samples were collected for 1 year and for enterovirus concentration the adsorption-elution on nitrocellulose membrane was used and for HEV, two methods of virus concentration were used, urea arginine phosphate buffer [U-APB] and PEG 8000.The results of HEV investigation of aborted women sera was 22% for IgG, 3% for IgM, 20% HEV-Ag, and 16% of HEV-RNA by RT-PCR. For fetal tissue, HEV-Ag was detected in 5% of the collected samples. The detected enteroviruses were coxsackieviruses types 2, 3,4and 5 in all serum samples and wastewater samples. The results showed also, that virus concentration by U-APB is much better than PEG-8000 but not highly efficient

Examination for Hepatitis E virus in wastewater treatment plants and workers by nested RT-PCR and ELISA

Water and sera were collected from four sewage treatment plants whose workers were exposed to raw untreated sewage samples which may accumulate the HEV virus. Two methods of water concentration were used [PEG and U-APB] and the samples were analyzed by nested RT-PCR. The 24 samples [inlet and outlet] of sewage treatment plants were negative for the virus. HEV IgG was detected in 205 workers sera and the age of 20-40 years exhibited the highest HEV IgG infection [50.9%- 50.4]. The highest infections between workers were in Balaks Sewage Treatment Plant and the persistence of HEV IgG was until 60 years old

Detection of hepatitis E virus in greater Cairo two wastewater treatment plants and its prevalence among workers of these plants

Because of inadequate public sanitation, epidemics of HEV infection have been reported in several developing countries. HEV-specific cDNA was prepared by reverse transcription of the total RNA extracted from water samples. Specified DNA amplification by PCR demonstrates the presence of HEV in sewage samples from the inlets [PCR positive 10/36:27.77%], while outlet samples were PCR negative. HEV IgM was detected in 40 workers out of 78 in these two studied plants [age 20-60 years], with a percentage of 51.25% and HAV IgM was also detected in 3 workers out of 78 [3.84%]. The study of serum ALT and albumin level go with the increase in HEV IgM in sera. This study which was carried out in two different wastewater treatment plants showed that HEV contamination was higher in one of them [El-Berka] than El-Zenin. A total of 33.33% of influent water samples were positive and 55.25% of workers were HEV IgM positive in the first plant [E1-Berka] and the corresponding figure in the other plant [El-Zenin] showed lower contamination, 22.22% in influent water sample and 40.9% of workers