Molecular evaluation of M2 protein of Iranian avian influenza viruses of H9N2 subtype in order to find mutations of adamantane drug resistance

Background: the H9N2 subtype of influenza A viruses is considered to be widespread in poultry industry. Adamantane is a group of antiviral agents which is effective both in prevention and treatment of influenza A virus infections. These drugs inhibit M2 protein ion channel which has role on viral replication

Objectives: the main objective of this study is to evaluate M gene of avian influenza viruses [AIVs] of H9N2 subtype in order to find adamantane drug resistance mutations

Methods: over 100 suspected samples were collected from different geographical regions of Iran during 2012-2013. Samples were injected via allantoic sac of 9-11 day-old chicken embryos. A total of 11 out of 100 were AIV. The H9N2 subtype was confirmed by specific RT-PCR. The RT-PCR was conducted for full length M gene. PCR amplified products were purified and then conducted for commercial direct sequencing. Finally, sequences were checked for possible sites of adamantane resistance mutations

Results: overall, 8 out of 11 viruses harbored the adamantine resistance-associated mutations. Of which, four viruses were isolated in 2013 and four viruses in 2012. Two different resistance- associated mutations were observed during different years

Conclusions: the present study provided clear evidence concerning resistance AIVs of H9N2 subtype that were circulating in Iranian poultry sector. This concern is always present as M segment might be introduced into human influenza viruses by reassortment phenomenon

Estimation of influenza and severe acute respiratory illness incidence [burden] in three provinces of the Islamic Republic of Iran, 2012 and 2013

There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection [SARI] and influenza-associated SARI [F-SARI] in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza [F-SARI] over 12 months [seasonality], and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend [seasonality] of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years

Measles vaccination coverage and seroprevalence of anti-measles antibody in south-east Islamic Republic of Iran

Discrepancies often exist between recorded immunization coverage and the real immunity level in a community. To estimate the vaccination coverage against measles in south-east Islamic Republic of Iran, a cross-sectional study was conducted in 3 districts during summer 2011. Using probability proportional to size cluster sampling, 1368 children aged 30-54 months were selected. Serum samples of 663 who had received 2 injections of mumps-measles-rubella [MMR] vaccine were checked for anti-measles IgG. Vaccination coverage for the second dose of MMR vaccine was 93.7%. The prevalence of anti-measles IgG in those who had received at least 2 MMR vaccine doses was 94.6%. There was a statistically significant association between the serological results and variables that reflected poor accessibility to health services. Combining serological results with coverage data, the proportion of the community protected against measles was estimated as 88.6%, which was below the limits defined for the measles elimination goals

comparative analysis of routine techniques: Reverse transcriptase polymerase chain reaction [RT-PCR] and five cell lines for detection of enteroviruses in stool specimens

Each year, Enteroviruses infect millions of people and cause different diseases. The agents are usually detected using cell culture. RD [Rhabdomyosarcoma] and L20B [L cells] are among the recommended cells by the World Health Organisation [WHO] for this purpose. Even though cell culture is the most common method used in diagnosing Enteroviruses in stool specimens, this particular method poses some problems, which include false positive or negative results, lack of a unique cell line for diagnosing all Entero virus types in addition to being time consuming. For these reasons, an attempt was made to find better techniques of Entero virus detection. RT-PCR [Reverse Transcriptase Polymerase Chain Reaction] is a technique used in place of the cell culture method. In this study, the cell culture method was compared with RT-PCR for detection of Enteroviruses in stool specimens. First, the chloroform treated stool samples were inoculated onto five cell lines, including RD, L20B, Hep-2 [Human Epidermoid carcinoma cell line], Vero [Verde Reno] and GMK [Green Monkey Kidney]. The results were then compared with data from Entero virus detection using the RT-PCR technique. The difference between RT-PCR and cell culture results was significant. Enteroviruses were detected in 24% of specimens using RT-PCR while cell lines could isolate Enteroviruses in just 14.4% of the samples

Molecular epidemiology of measles virus before and after the 2003 mass vaccination campaign for measles/rubella in Iran

Molecular epidemiology of measles virus [MV] is important, not only to measure the success of measles vaccination programs but also to monitor the circulation and elimination of the virus worldwide. In this study, we compared MV obtained from patients before the 2003 mass vaccination MR campaign and viruses detected after 2003 until 2008 in Iran. The nucleoprotein [N] gene of 29 MV strains circulating in Iran between 2002 and 2008 were amplified by RT-PCR and subjected to sequence and phylogenetic analysis. Molecular characterization of MV studied here revealed that although the outbreaks in Iran were associated with MV genotype D4, the isolated viruses clearly belonged to several different lineages. Maximum and minimum homology within the 29 Iranian strains in our study was100% and 94.9% within the carboxyl terminus of the N gene, respectively. Using Clustal X program, the alignment of Iranian MV sequences showed nine lineages. This study provides the usefulness of MV sequence analysis for the demonstration of local interruption of indigenous strain transmission as well as providing a valuable means for monitoring the elimination processes of MV control

First report of respiratory syncytial virus and human metapneumovirus co-infection in a 2-year-old Kawasaki patient in Iran

Respiratory virus infections in children are a leading cause of morbidity and mortality worldwide. A total of 897 clinical specimens were collected from February 2007 to January 2008 and transported to the National Influenza Center. Two hundred and two samples belonged to children under the age of six from 897 specimens, described above, were selected. Then they were tested for influenza virus types and subtypes by real time PCR assay subsequently, the specimens were tested for RSV and hMPV by hemi-nested multiplex PCR and parainfluenza viruses type 1-4 by hemi-nested mutiplex PCR and adenovirus by hemi-nested PCR. The throat swab was taken from the Kawasaki case with the history of chicken's contact. The specimen was tested for all influenza subtypes especially H5N1 and the results were negative. Meanwhile PCR was done for screening of other respiratory viruses that results came out positive for RSV and hMPV. In the present study, we demonstrated the possibility to detect dual infection caused by RSV and hMPV, but because of the extravagant pattern of this case, more investigation is suggested specially on Kawasaki patients

[Prevalence of anti-HCV antibody and related risk factors among bleeding disorder patients in Yazd province of Iran]

Inherited bleeding disorder is a disease due to deficiency in clotting factors or platelets. Replacement therapy of unscreened blood and blood products or unviricidal concentrated clotting factors would expose these patients to risk of acquired hepatitis C. The aim of this study was to determine the seroprevalence of hepatitis C virus and related risk factors among bleeding disorder patients in Yazd province. This cross-sectional study was performed in Yazd province in summer 2006 using census method. Following data collection through questionnaires, blood samples were taken from 77 [82.8%] patients. Plasma samples were tested for the presence of anti-HCV antibody via a third-generation ELISA kit. To exclude false positive cases, ELISA positive samples were re-tested by the confirmatory third generation RIBA test. The assessment of risk factors was done from the information analysis of both the questionnaires and test results by Chi-square test and multivariate logistic regression analysis, using SPSS 11.5 statistic software. The frequency of anti-HCV antibody was found in 38 [49.4%] patients. Chi-square test showed a statistically significant relationship between the severe form of the disease [in terms of blood products needed] [P<0.001], treatment duration of more than 121 months [P<0.001] and family history to HCV [P<0.05] antibody positivity. Successful execution of the screening of the blood and blood products as well as more scrutiny in preparing concentrated factors have been led to a remarkable decrease in the new infections among the recipients of these products in Yazd province

[Neutralization of helicobacter pylori cytotoxicity on vero cells by omeprazole]

Omeprazole is a gastric parietal cells proton pump inhibitor that is also active against H. pylori in vitro. This study was designed to examine the neutralization of H. pylori cytotoxicity on Vero cells by omeprazole micronized in strains isolated from gastritis, ulcer, cancer and Barrett's ulcer, to determine whether omeprazole can inhibit vacuolation of the Vero cells induced by cytotoxin of H. pylori or by urease. The effect of omeprazole on motility of H. pylori was assessed using concentrations lower than MIC. The antimicrobial activity of omeprazole micronized was studied by determining the MICs for 15 H. pylori strains. Water extract of the bacteria [concentrated culture supernatant] and different concentrations of omeprazole were added to Vero cells in culture. Also extracted urease from H. pylori strains with urea [10 mM] and omeprazole were added to Vero cells in culture. The inhibitory effect of omeprazole on motility of H. pylori was tested in semi-solid medium. MIC of omeprazole micronized was 20 micro g/ml. Omeprazole could inhibit induced vacuolation by the water extract of H. pylori strains in Vero cells. It could also inhibit vacuolation induced by urease. Inhibition of vacuolation strains was assessed microscopically and by the neutral red method. It was also found that omeprazole inhibits the motility of H. pylori strains at concentrations lower than MIC. The results of this study suggest that omeprazole micronized could neutralize the vacuolation effect of H. pylori cytotoxin on Vero cells probably by targeting v-type ATPase. The bacterial motility was also inhibited by low concentrations of omeprazole. The results of this study considers omeprazole micronized as an effective drug which targets important virulence factors of H. pylori including vacuolating cytotoxin, urease, and motility

Influenza surveillance in the Islamic Republic of Iran from 1991 to 2001

To better underst and the annual distribution of influenza virus in our country, we isolated and typed 45 viruses from 1043 patients with acute respiratory illnesses in a 10-year study conducted by the National Influenza Centre of the Islamic Republic of Iran. The seasonal distribution of influenza typically ran from November to April. Type A influenza was most common during the winters of 1991-92, 1997-98 and 2000-01 and type B influenza was most common during 1992-5 and 1996-97. Both type A and type B viruses circulated in 1995-96 and 1998-2000. A serological survey based on haemagglutination inhibition test confirmed our findings. The annual pattern of strains isolated was similar to the worldwide pattern during the same interval

[Epidemiologic survey of documented measles outbreak in Tehran]

Background: in spite of extended vaccination programs against measles, it is the most preventive contagious disease that leads to death. For adaptation of WHO policy for measles in Iran; we should determine susceptible age population whom at present lay out of current vaccination program

Methods and Materials: in this descriptive study which was performed in the public health School of the Tehran medical university we obtained 13 I documented measles cases among 650 suspected case of measles [as WHO discrimination] According to serum samples analysis [ capture IgM EIA 71% and 29 % HJ]

Results: in this study 63. 4 % of cases were male and 39.6 % were females. Between patients, the most prevalent age group was between 15 - 19 years old. Two peaks of the time of the disease were March and June. History of past vaccination against measles showed that 43 % had unknown history; 24. 4 % had two vaccinations 39. 3 % had one vaccination and 3.1 % had no vaccination history

Conclusions: it is suggested [by further investigations in future] the need for revaccination of measles/or age group of 15-19 in Iran will be determined

Prevalence of human papillomavirus in Mazandaran Province, Islamic Republic of Iran

We report the prevalence of human papillomavirus [HPV] types in 100 cervical biopsy specimens in Mazandaran province. HPV DNA was detected in 78.6% of cervical carcinoma cases, 64.3% of dys/ metaplasia and 9% of normal cases. Significant correlation was found between the presence of HPV DNA and development of cervical carcinoma. HPV types 16 and 18 were detected in 60.6% of HPV-positive cervical carcinoma cases, whereas HPV31 and 33 were found in 21.2%, and HPV6 and 11 in 18.2%. Among HPV-positive dys/metaplasia cases, 55.6% were positive for HPV16 and 18, 22.3% for HPV6 and 11, and 11.1% for HPV31 and 33. Only HPV6 and 11 were detected in 4 [100%] normal biopsy specimens