Detection of rtN236T mutation associated with adefovir dipivoxil resistance in Hepatitis B infected patients with YMDD mutations in Tehran

Background Objectives: The risk of adefovir dipivoxil resistance emergence has increased in lamivudine-resistant hepatitis B infected patients. The mutations known as causing adefovir resistance, rtN236T and rtA181V/T, are detected within the D and B functional domain of the HBV polymerase, respectively. In this study, we intended to determine the pre-existing adefovir-resistance mutations in patients infected with LAM resistant mutants prior to starting adefovir therapy. Material and Methods: The study included 30 patients with chronic hepatitis B with lamivudine resistance mutations in the YMDD motif that experienced viral breakthrough. Results: After alignment of protein coding sequences, the rtN236T mutation was observed in two (6.6 %) patients, while twenty-eight others had neither rtN236T, nor rtA181V/T mutation. All 30 patients were infected with genotype D of hepatitis B virus. Conclusions: The early detection of LAM-resistance mutations may allow a timely chance of therapy to avoid hepatitis flare-up. This data suggests that monitoring of ADV-resistance mutations in ADV naïve patients can be considered in selecting the appropriate anti-viral regimen

Human Bocavirus in Iranian children with acute gastroenteritis

Background: Human Bocavirus (HBoV) infection is of worldwide distribution. There is increasing evidence that HBoV is pathogenic for the human gastroenteric tract. However, less data are available on the role of HBoV in gastroenteritis. The present study was aimed to determine the prevalence of HBoV in children with gastroenteritis. Methods: Real-time PCR TaqMan was used to screen 200 stool specimens that had been referred to the virology laboratory for HBoV evaluation. All of samples were collected on viral transport media. Results: Of the 200 stool samples analyzed, 16 (8%) were positive for HBoV. Human Bocavirus positive samples from patients aged between 1 to 5 years with acute gastroenteritis infection suggest a minor role of HBoV in gastroenteritis (p=0.0001). Conclusion: The study showed a high prevalence of human Bocavirus in young children with acute gastroenteritis diseases in Iran, suggesting that HBoV play a role in the pathogenesis of gastroenteritis. Keywords: Gastroenteritis, Child, Human Bocavirus, Real-time PCR

Relation between parvovirus B19 infection and fetal mortality and spontaneous abortion

Background: Infection with parvovirus B19 may cause fetal losses including spontaneous abortion, intrauterine fetal death and non-immune hydrops fetalis. The aim of this study is to determine the frequency of parvovirus B19 in formalin fixed placental tissues in lost fetuses using real-time PCR method. Methods: In this cross-sectional study, 100 formalin fixed placental tissues with unknown cause of fetal death were determined using real-time PCR method after DNA extraction. Results: Six out of 100 cases (6%) were positive for parvovirus B19 using real-time PCR. Gestational age of all positive cases was less than 20 weeks with a mean of 12.3 weeks. Three cases have a history of abortion and all of positive cases were collected in spring. Mean age of positive cases were 28 years. Conclusion: Parvovirus B19 during pregnancy can infect red precursor cells and induces apoptosis or lyses these cells that resulting in anemia and congestive heart failure leading to fetal death. Management of parvovirus B19 infection in pregnant women is important because immediate diagnosis and transfusion in hydropsic fetuses can decrease the risk of fetal death

Human Herpesvirus-6 and Human Herpesvirus-7 Infection in Iranian Patients with Neurological Illness

Human herpes virus-6 (HHV-6) and HHV-7 have been implicated as causes of meningitis and encephalitis in children and adults. In this study the presence HHV-6 and HHV-7 DNA were tested in cerebrospinal fluid (CSF) sample taken from Iranian children, suffered from meningoencephalitis. From 2007 to 2009, 150 patients from Tehran with meningoencephalits who were referred to a pediatric ward in Rasoul Akram hospital, Tehran Iran, were enrolled in the present study. Conventional and BACTEC Ped Plus medium were used in conjunction with latex agglutination test and real time PCR for detection of HHV-6 and HHV-7 DNA in clinical specimen. All type of human herpes virus DNA was detected in 12 % (18/150) cases. HHV-6 DNA was detected in 4.7% and HHV-7 DNA was detected in 2 cases (1.4%). Human herpes virus-6 and HHV-7 DNA was detected in 6% of all studied cases. HHV-6 was slightly more frequent than HHV-7. Our findings were lower than the rate of other references but were higher than the findings of previous study in Iran. This variation might be due to differences in methods, age of study cases or epidemiologic and geographic variation

Congenital rubella infection in neonatal cord blood samples of newborns in hospitals affiliated to Tehran University of Medical Sciences

Rubella is a disease caused by the rubella virus and it is usually mild and self-limiting. Infection of a developing fetus is serious and important because the child may be born with congenital rubella syndrome. Its symptoms include mental retardation, heart defects, cataract, etc. In 2003, mass vaccination against measles and rubella in individuals 5-25 years old was done. One of the main objectives of this study was to survey congenital rubella infection status with the presence of IgM antibodies against rubella virus in cord blood samples and also the immunity assessment of maternal IgG antibodies against rubella virus in the above samples. Methods: The cross-sectional study was to determine the transfer of congenital rubella in 358 cord blood samples collected in hospitals affiliated to the Tehran University of Medical Sciences that was done in 2008-2009 The collected samples were analyzed by two ELISA methods for detection of IgG and IgM antibodies, RT-Nested PCR tests was applied on samples of IgG–negative and IgM-positive and also some of randomly IgG-positive samples for identifying the presence of the virus genome. In this study two groups of mothers were tested, one consisted above 29 years of age (at the time of vaccination) with the frequency of 73.4% and the other one below 29 years of age with the frequency of 26.6%.Results: Of the 358 samples, 91.1% IgG and 2.8% were found to be positive. None of the 31 samples were positive according to the presence of the virus genome via the method of RT-Nested PCR. Conclusion: According to high immunity of mothers, the probability of congenital rubella transmission was low, but because of low immunity of mothers of >29 years of age, it is much better to upgrade the age of vaccination to 28 years old

Antiviral Activity of Sirna UL42 against Herpes Simplex Virus Type 1 in HeLa Cell Culture

RNA interference (RNAi) is a process by which introduced small interfering RNA (siRNA) can cause the specific degradation of mRNA with identical sequences. In this study, we applied siRNAs targeting the UL42 gene of human herpes simplex virus type 1 (HSV-1), which encodes a multifunctional polypeptide that is vital for virus DNA replication, binding to DNA, stably associating with the virus DNA polymerase (Pol), and acting to increase the length of DNA chains synthesized by Pol. HeLa cell line was used for HSV 1 infection and SiRNA transfection was done to suppress UL-42 gene in cell culture. The decrease in titer of HSV 1 was observed by rReal Time PCR to detect the drop in HSV 1 DNA synthesis and translation. The inhibition rates of siRNA1 and siRNA2 on HSV-1 plaque formation were reported and Comparing with virus control, siRNA1 and siRNA2 reduced DNA replication HSV-1. The decision whether the decrease in the number of HSV-1 plaques was due to siRNA silencing expression of the UL42 gene, a real-time PCR indicating that UL42-specific siRNAs blocked the expression of the UL42 gene significantly. Comparing with virus control, siRNA1 and siRNA2 reduced the expression of UL42 gene. In this study the synthetic siRNA silenced UL42 mRNA expression effectively and specifically and inhibited HSV-1 replication and also our data offer new possibilities for RNAi as a genetic tool for inhibition of HSV-1 replication

Detection of cytomegalovirus (CMV) antibodies or DNA sequences from ostensibly healthy Iranian mothers and their neonates

Cytomegalovirus (CMV) remains the most common cause of viral intrauterine infection. The objective of this research was to determine the prevalence of at-risk pregnancies for congenital cytomegalovirus transmission in a randomly selected pregnant women and their newborns. Enzyme Link Immunosorbent Assay (ELISA) and real-time polymerase chain reaction (PCR) were utilized to screen the sera of mothers (n = 100) and consecutive umbilical cord blood samples from their newborn (n = 100). Of the 100 mother's sera analyzed, 100 (100%) and 3 (3%) were positive for cytomegalovirus IgG and IgM antibodies, respectively. Of the 100 cord serum specimens analyzed, 99 (99%) and 2 (2%) were positive for cytomegalovirus IgG and IgM antibodies, respectively. Cytomegalovirus DNA was detected in 4 out of 100 (4%) cord blood samples of newborns. From four CMV DNA positive cases, Case 1 had no IgM in cord serum, but had IgM in mother's sera. Cases 2 and 4 were positive for IgM in both mother's sera and cord serum. Case 3 had no detectable CMV IgM in sera and cord serum. As many as 66 and 100% of CMV IgM-positive women in this study also had CMV IgM and CMV DNA in their delivery cord blood samples, respectively suggesting an increased risk of congenital CMV infection in those pregnancies. A paired women sera/cord blood CMV IgM-negative was found to be positive for CMV DNA. The data may also suggest the utility of PCR in place of CMV IgM as a diagnostic method for congenital CMV infection

Detection of human T-cell lymphotropic virus Type-1 among patients with malignant hematological diseases in Capital of Iran, Tehran

Human T-cell lymphotropic virus type-1 (HTLV-1) is a deltaretrovirus linked causally to adult T-cell leukemia or lymphoma (ATL), and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The aim of this study was to detect HTLV-1 infection in patients with malignant hematological diseases and also determining the prevalence of HTLV-1 in these patient groups. Sixty patients with malignant hematological diseases were included in the study and tested by enzyme-linked immunosorbent assay (ELISA) for anti-HTLV-1, and Real time-PCR for the sequences from HTLV-1 tax gene. The mean age of patients was 33.9 ± 18.3 years. 18 of the subjects were found HTLV-1 seropositive using ELISA and the viral prevalence by Real time-PCR was 12%. HTLV-1 was found in 25% of patients with acute myelogenous leukemia (AML), 58.3% of patients with chronic myelogenous leukemia (CML), 16.7% of patients with acute lymphoblastic leukemia (ALL), and no detected in patients with lymphoma. The present study revealed that HTLV-1 is prevalent in patients with malignant hematological diseases and in our study. The major HTLV-1 associated syndromes were chronic myelogenous leukemia and acute lymphoblastic leukemia

Middle East Respiratory Syndrome Coronavirus (MERS-CoV)

The Middle East respiratory syndrome coronavirus (MERS-CoV) as a causative agent of severe lower respiratory tract infection in humans, considered as a global threat, especially against to Persian Gulf countries. Since its discovery in 2012, MERS-CoV has spread 27 countries affecting about 1800 people and caused more than 600 deaths in worldwide. In comparison to SARS (severe acute respiratory syndrome), MERS-CoV appears to have a higher mortality rate (40% versus 10%) and is particularly more severe in patients with underlying medical conditions. Until now, the most MERS-CoV cases (more than 85 percent) have had a history of travel or residence in the Middle East countries. A possible intermediate host for MERS-CoV is camel. Clinical manifestations of MERS range from mild or asymptomatic disease to acute respiratory syndrome and multi-organ failure resulting in death, mostly in individuals with preexisting medical co-morbidities. There is no specific antiviral treatment for MERS and infection prevention and control practices are necessary to prevent spread of MERS-CoV in health care facilities. In present study, we have briefly outlined the recent information about the epidemiology, clinical features, diagnosis, treatment and prevention of MERS-CoV

Design of primers for pertussis diagnosis by Real Time PCR and determination of its sensitivity and specificity in comparison with commercial kits.

Background and Aim: Pertussis vaccination in this country has been going on for many years and active infection or vaccination will provide immunity in 85% of cases. However, every 2-5 years outbreaks in unprotected adults creates an epidemy for children and infants. Based on conserved genomic sequences, Real time PCR could be an easy, cost- benefit, fast and highly sensitive method for pertussis detection. Materials and Methods: A total of 170 nasopharyngeal swabs of infants with history of cough for more than 2 weeks were collected. In the first stage, Bordetella pertussis bacteria detection was performed by culture and followed by Real time PCR using a commercial kit and then repeated with newly designed primers. Results: Performance of our home made primers for detecting pertussis using Real Time PCR in comparison with those by commercial kit was acceptable based on diagnostic classical guidance (WHO) and the (CDC). Conclusions: Real time PCR test with new primers in comparison with culture techniques is more suitable, high sensitivity and can provide more informative values for pertussis detection