Showing 47 results for Virus
T Mokhtari Azad , H Mohammadi , M Mahmoodi , Z Saadatmand , A Moosavi , R Hamkar , R Nategh ,
Volume 57, Issue 2 (5-1999)
Abstract
This is a report of the first serological survey of influenza C virus in Iran, performed during a one year period (March 1997-May 1998). This study was accomplished in the National Influenza Center-Division of Virology in Tehran University of Medical Scinces. 1080 samples of serum (689 samples from Tehran and 391 samples from other provinces) were assayed for the presence of antibodies against influenza C virus (C/Paris/1/67) by haemagglutination inhibition (HI) test. 43.7% of people tested in Tehran and 40.7% of people tested from other provinces had protective antibodies against influenza C virus. Distribution of seropositives in various age groups had a somewhat similar pattern as what has been reported from other countries. The results of this study indicates that the lowest level of protective antibody titer is found at childhood and the level increases with age. The protective antibody titer level off for 20-30 years old age group and decreases in older age groups. These results indicates a primary contact in childhood, reinfection in adulthood. The influenza C virus is simultaneously circulating in Iran with other types of influenza viruses (types A and B).
E. Keyhani, N. Kohannia, N. Izadimood, M. R. Keyhkhaee, H. Najmabadi,
Volume 64, Issue 3 (5-2006)
Abstract
Background: Cervical cancer is the second leading cause of cancer death among women. In this cancer, the effects of prevention, early diagnosis and treatment more than other cancers decrease the mortality rate. In 1970 human papilloma virus (HPV) was introduction as major etiologic factor of cervical cancer. Different studies throughout the world revealed strong correlation between HPV and cancerous & precancerous changes in epithelial cells. Since cell culture and serological methods can not recognize the virus and its subtypes, the importance of the molecular methods including polymerase chain reaction (PCR) in early and definite diagnosis of virus is obvious.
Methods: In this study, after patient selection using the related protocol and completion of the questionnaires, 100 samples from cancer lesions of cervix selected. Then DNA extraction from paraffin blocks performed using standard method. Multiplex PCR with two pairs of primer (one as internal control) performed and the PCR product run on 8% polyacrylamid gel.
Results: The results showed that 73% of the tissues were infected by HPV.
Conclusion: This finding confirm the previous results based of correlation between HPV,and cervical cancer.
Yarandi F, Izadi Mood N, Eftekhar Z, Niakan R, Tajziachi S,
Volume 65, Issue 14 (3-2008)
Abstract
Background: Cervical cancer is the second most common cancer of the women
worldwide. It is also an important cause of cancer-related mortality in women, after breast
cancer. Nearly half million of new cases are identified yearly. The incidence rate in
developing countries is greater than the developed countries. Epidemiologic studies have
shown that the association of genital human papilloma virus (HPV) with cervical cancer is
strong, independent of other risk factors, and consistent in several countries. The aim of
this study was to determine the frequency of HPV in patients with high grade cervical
intraepithelial neoplasia (CINIII, CIN II) and squamous cell carcinoma (SCC) of cervix.
Methods: Hundred specimens from patients with SCC and CINIII, CIN II, confirmed by
histological review, referring to Mirza Koochak Khan Hospital from 1999-2004 were
enrolled in a cross sectional study. Polymerase chain reaction was utilized for identification
and typing of HPV DNA. To increase the sensitivity of HPV detection, nested PCRs were
performed using MY09/MY11 as outer and GP5/GP6 as inner primers.
Results: It was possible to extract 77 of 100 specimens that HPV DNA was detected in 47 of
77 specimens. Infection with HPV was present in 32 specimens (86.5%) among SCC patients
and in 15 specimens (37.5%) among CINIII, CIN II patients. The most frequent HPV types in
SCC patients were HPV 16 and 18 (59.38%) and then 33 (34.38%) and in CINIII, CIN II
patients was 16 (53.33%) and 18 (40%). the most frequent co-infection in both groups was
HPV 16 and 18 which was present in 40.62% and 26.7% of cases respectively.
Conclusions: The most frequent HPV types in patients with SCC and CINIII, CIN II
was 16 and 18 that is identical to many other countries infection pattern.
Noorbakhsh S, Tabatabaei A, Parvaresh M, Tonekaboni H,
Volume 66, Issue 10 (1-2009)
Abstract
Background: In recent years, many ill cases with cytomegalovirus reactivation in non-immuno compromised persons reported. Goal of study: to determine the CMV infection in cerebrospinal fluid of aseptic meningoencephalitis children hospitalized in Rasul & Mofid hospital (2005-2007).
Methods: In a cross sectional study 132 cases selected with simple sampling. CMV-DNA in their Cerebro spinal fluids searched by qualitative PCR.
Results: The age range of the study patients was 5 month- 13 years, median age= 2±3.7 years 87(65.9%) male and 45(34.1%) was female. The presenting signs and symptoms were convulsion 77(69.4%) meningitis 25(18.8%), loss of consciousness 47(37%) neurologic defects 15.9%. DNA extrated in 11 cases. Mycoplasma- DNA in 2cases DNA-CMV detected 2(1.5%). Positive DNA HSV found in 7(15.3%) of patients. DNA- HSV type- 15.3% (7/132) cases. An infant 5 month age with developmental delay, microcephaly and recurrent convulsions. A 1 year girl with brain atrophy and progressive hydrocephaly with intracranial shunt
Conclusions: Differentiation between herpes meningoencephalitis and other encephalopathy based on clinical signs in children is too difficult. CMV (1.5%) has lower rate than herpes simplex type-1 (5.7%). In addition to CMV and HSV1 all of herpes family viruses (varicella, herpes 6, 7, Epstein barr virus) could have role in children with meningoencephalitis. In recent years a sensitive, rapid, simple diagnostic test "Single tube Multiplex PCR" in cerebro spinal fluid recommend. Rapid diagnosis and faster treatment is necessary for decreasing mortality and morbidity in all of herpes meningoencephalitis cases
Jafari S, Rasoolinejad M, Emadi Kouchak H, Mokarami F,
Volume 67, Issue 7 (10-2009)
Abstract
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Background: CD4 T-Lymphocyte counts have proven to be a standard
laboratory marker of disease progression and severity of immunodeficiency in
adults infected with HIV is used to initiate and monitor highly active
antiretroviral therapy however, its application may not be feasible for its
expensive equipments and reagent in resource-limited setting. There is a need
to have another marker of immunodeficiency that is less resource-demanding. In
April 2002, the World Health Organization (WHO) recommended that, when CD4 cell count is not available, a
TLC of
1200cell/mm3
or less in individuals with stage 2 or 3 of the disease may be used as an indication to
initiate ART.
Methods: The aim of this study was to determine the
relationship between total lymphocyte count and CD4 count in HIV-infected
adults. This was a retrospective cross-sectional study. Subject characteristics
were patients who had positive serologic HIV test results, confirmed via western blot. Analysis unit was the
results of CBC and CD4 measurements on the same blood sample each time. Data
of 100
patients were collected. In this study, TLC accounts for the main predictor of CD4 count. The
amounts of TLC which can predict CD4 less than 200cell/mm3 were considered eligible.
Results: Our data revealed high sensitivity and specificity of TLC as a surrogate
measure of CD4 count. In this study, TLC cutoff of 1300cell/mm3 indicated the optimal combined sensitivity and specificity altogether.
Conclusion: Total lymphocyte count and its changes can be used as
alternative to CD4
count and its changes in the management of HIV-infected individuals.
Hadadi A, Moradmand Badie S, Roham M, Rasulinejad M, Mirzai N,
Volume 67, Issue 8 (11-2009)
Abstract
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Background: One of the clinical manifestations of Human Immunodeficiency Virus (HIV)
infected patients is cardiovascular disorder. The aim of this study was to
evaluate the prevalence of cardiovascular disorders in HIV
infected patients for the beginning treatment of these patients and reducing
mortality and morbidity in these patients.
Methods: This cross-sectional study was performed on 134 HIV
infected patients who referred to Imam Khomeini hospital, Tehran University of
Medical sciences, Tehran Iran during years 2007-2008.
Demographic characteristics, history of smoking and opium addiction,
antiretroviral therapy, class of drugs and duration of consumption were
recorded. After completion of physical examination, electrocardiography and
echocardiography studies were done.
Results: In this study 98(73.1%) patients were male.
The mean age of the patients was 36.5±10.3
years. The mean of the CD4 number were 296±181. Injection drug users were 54.4%
of the study patients. Cardiovascular disorders were found in 84(62.7%)
patients. Among patients with heart diseases, 75%
were male. The most Electrocardiographic change was the axis deviation of the
heart found in 32(23.7%) patients.
Pericardial effusion and LVEF<50% were
noted in 7(5.2%) and 23(17.2%)
patients respectively. The involvement of the mitral valve
in 59(44%), tricuspid valve in 21(15.7%)
and aortic valve in 6(4.5%) patients were
noted. Myocardial dysfunctions existed in 10(7.4%)
patients.
Conclusions: Our results showed a high prevalence of cardiovascular disorder in HIV
infected patients. We recommend the evaluation of the cardiovascular
system in all HIV infected patients
even if they are symptom free.
Pourakbari B, Mamishi S, Pajand O, Nadji Sar, Mahjob F, Kochakzadeh L, Izadyar M, Parvaneh N, Saboni F,
Volume 67, Issue 11 (2-2010)
Abstract
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Background: Latent Epstein- Barr
virus (EBV) genomes are found in the malignant cells of
approximately one-third of Hodgkin's lymphoma (HL) cases. Detection of EBV viral DNA could potentially be used as a biomarker of disease activity. Our goal was
to compare of EBV DNA detection in samples obtained from lymphoma
patients versus controls.
Methods: One milliliter uncoagulated and 1ml coagulated blood sample for DNA extraction and serum analysis using ELISA for IgG anti EBNA-1 were obtained from 44 lymphoma patients and from 44 normal controls,
respectively. EBV genome, EBNA-2, was examined from DNA extracts of paraffin
embedded and blood samples using Nested PCR with type specific inner primers.
Results: Positive results for ELISA, Blood and biopsy PCR in study group were, 84.1%, 27.3% and 13.6%, respectively. However, these results in
control group were 47.7% and 16% for ELISA and Blood PCR assays, respectively. Positive results in ELISA, Blood PCR and Biopsy PCR in Hodgkin and non-Hodgkin patients were found in 21(84%), 6(24%), 4(16%) and 16(84.2%), 6(31.6%), 2(10.5%) of specimens, respectively. No significant
differences in EBV detection were found between these two
patient groups (p values for ELISA, Blood PCR and Biopsy PCR were 0.26, 0.73 and 0.68, respectively).
Conclusion: Comparison of ELISA and Blood PCR results in children and adult
patients with the same age of controls have showed difference in ELISA results of children, only.
None of the test results have showed statistically significant difference
between Hodgkin and non-Hodgkin patients. However, the mean of ELISA results in Hodgkin patients
was higher as compared with controls. Blood PCR assay cannot be recommended as a biomarker of disease activity in EBV positive Hodgkin's lymphoma
patients.
Mousavi A, Akhavan S,
Volume 68, Issue 2 (5-2010)
Abstract
Background: Primary clear cell adenocarcinoma of cervix (CCAC) is usually seen in women with a history of in utero exposure to diethyl acetyl bestrol (DES). We report two cases of clear cell adenocarcinoma of cervix with no history of exposure to DES in embryonic period.
Case presentation: The first case was a 14-year-old women with complaint of painless vaginal bleeding. There was atypical cells in Pap Smear and a bleeding tumor with 1.5 cm in diameter was found in vagina. She was admitted with a diagnosis of CCAC of the uterine cervix stage Ib2 according to FIGO classification.
The second case was a 23-year-old patient with complaint of painless vaginal bleeding. The results of cervical cytology was normal. Evaluation of the punch biopsy sample revealed CCAC. Her clinical exam showed stage IIb according to FIGO classification. Both patients had no history of exposure to DES during embryonic period. The first patient treated with radical abdominal hysterectomy and systematic pelvic lymphadenectomy and for the another one external beam radiotherapy and brachytherapy was performed. There was no any recurrence or metastasis after an 18-24 months follow-up
Conclusions: Primary clear cell carcinoma of cervix could be unrelated to HPV infection or exposure to DES during embryonic period and in approach to these patients
this subject should be considered.
Faghihloo E, Rezaie F, Salimi V, Naseri M, Mamishi S, Mahmoodi M, Mokhtari-Azad T,
Volume 68, Issue 3 (6-2010)
Abstract
Background: Human respiratory syncytial virus (HRSV) is the most important viral agent of acute lower respiratory tract disease in infants and young children worldwide. This virus is responsible for 50% brochiolitis and 25% pneumonia in infants. There are limited data of molecular epidemiology of HRSV from developing countries. This is the report on the molecular epidemiology of human respiratory syncytial virus in Iran. Methods: In this study, RT-PCR for second hypervariable region of the HRSV G glycoprotein was performed on 72 throat swabs collected from children less than 5 years of age with acute respiratory symptoms in 1386.
Results: Of the 72 throat swabs collected from children with acute respiratory symptoms, 14 (19.44%) were positive for HRSV. Phylogenetic analysis revealed that all HRSV-positive samples clustered in three genotypes of subgroup A: 12 strains (85/71%) in genotype GA2, 1 strain (7/1%) in genotype GA1, and 1 strain (7/1%) in genotype GA5. In this study we couldn’t identify any genotype of subgroup B.
Conclusion: Our results revealed that multiple genotypes of subgroup A were co- circulated during 1386 in children less than 5 years of age in Iran. Also this study revealed that genotype GA2 was predominant genotype in isolates were obtained from several cities (Tehran, Isfahan, Karaj, Qazvin, Bandar Abbas, Shahreza), so we speculate that this genotype may be predominant during 1386 in Iran. This study supported that RT-PCR for second variable region of G protein is an effective method for further studies of HRSV genotype designation in Iran.
Ahmadinejad Z, Soleimani Ar,
Volume 68, Issue 3 (6-2010)
Abstract
Background: Fever in neutropenic patients is a medical emergency which may happen in patients undergoing chemotherapy. The definition of neutropenia varies from institution to institution but is usually defined as an absolute neutrophil count (ANC)
< 500 cells/l or < 1,000 cells/l with a predicted nadir of < 500 cells/l. Bacterial and fungal infections are the most important in neutropenic patients. Viral infections with agents such as herpes simplex virus and cytomegalovirus are common but less than other pathogens.
Case presentation: We report a patient with fever & neutropenia following cytomegalovirus infection during chemotherapy course for breast cancer.
Conclusion: Although fever and neutropenia after cytomegalovirus infection is not very common but prompt diagnosis and treatment of this disease reduces the mortality and morbidity associated with cytomegalovirus. For this reason, screen testing for CMV
infection in high risk patients including patients with cancer and preemptive therapy in patients with viremia, for prevention of CMV disease could be considered as a strategy for prevention of CMV infection.
Shatizadeh Malekshahi S, Yavarian J, Naseri M, Rezaei F, Mokhtari Azad T,
Volume 68, Issue 9 (12-2010)
Abstract
Background: Respiratory virus infections represent a major public
health problem because of their worldwide occurrence, ease of spread in the
community and considerable morbidity and mortality. They are one of the most
common reasons for hospitalization of children under the age of six. In some
cases, infection with two different viruses increase the severity of disease
which lead to the hospitalization.
Case presentation: Among 202 samples related to children under the age of six
with respiratory infections, two dual infections of Adenovirus with other
respiratory viruses with PCR test were
detected.
Conclusion: Mixed respiratory viral infections are sometimes
associated with severe disease and recognition of coinfection is important. Dual infections of Adenovirus with respiratory syncytial
virus (RSV) and
Swine origin influenza A (H1N1) virus
were demonstrated. The evidence showed that the co-infection of Adenovirus with
swine origin influenza A (H1N1), has
increased the severity of disease which lead to the hospitalization.
Rahbarimanesh A, Saberi H, Modarres Gilani Sh, Salamati P, Akhtarkhavari H, Haghshenas Z,
Volume 69, Issue 8 (11-2011)
Abstract
Background: Acute gastroenteritis is a major cause of
morbidity and mortality among children in developing countries.
Rotaviruses are recognized as the most common etiologic factors of
gastroenteritis. In this study, we determined the epidemiologic
features, clinical symptoms and molecular structure of rotavirus VP4(P)
genotypes in children with acute diarrhea in Bahrami Hospital in Tehran
Iran, during 2009 for justifying the routine use of rotavirus vaccines
in children.
Methods: One hundred fifty fecal samples from
150 children with acute diarrhea in Bahrami Pediatric Hospital in
Tehran, Iran were collected from January to December 2009. The patients’
mean age was 20.90+18.19 years (ranging from 1 month to 14 years).
Fecal samples were transported on ice to the laboratory of virology
department of Pasture Institute of Iran. The demographic and clinical
data for each case were entered in an author-devised questionnaire.
Group A rotavirus was detected by dsRNA-PAGE. Subsequently, rotavirus
genotyping (VP4) was performed by semi-nested multiple RT-PCR and the
phylogenetic tree of the Rotavirus nucleotides was constructed. The data
were analyzed by statistical tests including Wilcoxon signed and
Mann-Whitney U.
Results: Rotavirus was isolated in 19.3% of the
samples, more than 90% of which had long RNA patterns. The predominant
genotype (VP4) was P[8] (86%) and other genotypes respectively were P[6]
(6.9%) and P[4] (6.9%).
Conclusion: A high prevalence of the P[8]
genotype was found to be the cause of acute diarrhea. The analysis of
P[8] genotype sequence showed a high level of similarity of the virus in
this study with those of other Asian countries.
Bokharaei-Salim F, Keyvani H, Zamani F, Jahanbakhsh Sefidi F, Amiri A,
Volume 69, Issue 10 (1-2012)
Abstract
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Background: Hepatitis
C
virus (HCV) is
essentially considered as hepatotropic, but virus sequences have also been
found in other important extrahepatic sites, including peripheral blood
mononuclear cells (PBMCs). This study was done to investigate the presence of mixed
infection and the differences between hepatitis C virus genotypes in plasma, peripheral blood mononuclear cells, and
liver biopsy specimens in patients with hepatitis C virus infection.
Methods : One hundred and fifty two patients with established chronic
hepatitis C infection attending Firouzgar Hospital, affiliated to Tehran
University of Medical Sciences, from September 2008
to April 2010 were enrolled in the present study. After collecting plasma,
peripheral blood mononuclear cell, and liver biopsy specimens, RNA was extracted from the samples
and hepatitis C virus genotyping was performed using INNO-LiPATM HCV II kit. The hepatitis C virus genotyping was confirmed by sequencing the RT-nested PCR product of 5'-UTR fragments.
Results : The mean age of the participants was 31.2±16.9
years. Multiple hepatitis C virus genotypes were detected
in 4 (2.6%) out
of 152 plasma
samples, 10 (6.6%) out of 152 peripheral blood mononuclear cell samples, and 9 (18.8%) out of 48 liver biopsy specimens. Hepatitis
C
virus genotypes were different in the plasma, PBMC, and liver biopsy specimens of 21
(13.8%) patients.
Conclusion: The present study
shows that a significant proportion of patients with chronic hepatitis C infection are infected by
multiple hepatitis C virus genotypes which may not be detectable in their plasma specimens.
Kosari F, Yarigarravesh Hr, Rezvan M,
Volume 70, Issue 6 (9-2012)
Abstract
Background: Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma. There are various types of DLBCL including immunoblastic and centroblastic. Epstein-Barr virus (EBV) is a member of Herpes virus family found in all human populations inducing different lymphoproliferative disorders. The role of EBV in the development of DLBCL is known. Multiple laboratory methods are available for detecting EBV. This study was conducted to determine the correlation of EBV with DLBCL in samples referred to pathology ward in Shariati and Sina Hospitals by chromogenic in situ hybridization (CISH) method.
Methods: In this case/control study, pathological specimens of 50 patients with DLBCL as well as 50 reactive lymph nodes and tonsils (control group) were collected from archives of Shariati and Sina Hospitals and were evaluated for EBV encoded RNA (EBER) expression based on CISH method. A peptide nucleic acid (PNA) EBV probe (Dakocytomatin) was used while all the processes were done in RNAase-free conditions using RNAase-free water, sterile gloves and samplers.
Results: Out of fifty specimens in the case group, eight were positive for EBER in comparison with two in the control group (P=0.046). No statistically significant difference was observed between intranodal or extranodal samples (P=0.736) or between males and females (P=0.0746).
Conclusion: Our study showed that EBV positivity for EBER in patient with DLBCL could be determined more effectively by CISH method than immunohistochemistry (IHC). Comparative analysis between CISH, PCR and IHC methods is recommended.
Reza Shah Siah, Mehran Sakaki, Zahra Safaie-Naraghi ,
Volume 71, Issue 7 (10-2013)
Abstract
Background: In the Mediterranean region , Kaposi's sarcoma (KS) has a high prevalence especially in patients with AIDS. Iran is located close to the Mediterranean region and the HIV prevalence is increasing in our country . In some stages, Kaposi's sarcoma is morphologically similar to other vascular tumors. Owing to the presence of human herpesvirus 8 (HHV-8) in all cases of Kaposi's sarcoma , detection of virus DNA by PCR method can help in the identification of non-diagnostic cases. Moreover, the prevalence of HHV-8 genotypes is different in various regions of the world and in different races. There are limited studies performed on the HHV-8 genotypes in Iranian population.
Methods: Patients with Kaposi's sarcoma from 2001 to 2011 who refer to Tehran Razi Hospital were enrolled in this study. HHV-8 DNA was extracted from paraffin blocks and amplification of the virus genome was performed by PCR method . Finally, the target DNA fragment was used for sequencing and genotype determination.
Results: PCR was performed on 53 cases. In 8 cases with suspicious morphology, PCR was negative and they were excluded from study. Of remaining 45 cases, 35 had positive PCR results, 7 had negative results and 3 had low PCR product. Samples from 28 cases that had positive PCR results, which were acceptable for genotyping, were chosen for sequencing. Twenty cases had genotype C, 7 cases had genotype A and one case was negative. The results are consistent with other studies in our geographical area. No correlation was found between the different microscopic stages and HHV-8 Genotypes.
Conclusion: Since the HHV-8 is obtained in almost 100% of KS lesions and PCR s ensitivity in detection of the virus is close to 100 %, KS diagnosis can be confirmed in suspicious cases by detection of HHV-8 DNA on paraffin blocks. Moreover the prevalence of HHV-8 genotype was determined in Iran.
Negar Sajjadian , Ramin Jahadi ,
Volume 71, Issue 9 (12-2013)
Abstract
Background: Anemia at the time of birth may cause some problem like asphyxia, heart failure shock or even death in a neonate. Different etiologies can be considered for this problem. Parvovirus B19, as a viral organism, can cause hydrops fetalis and neonatal anemia and consequent complications. We present here a case of newborn infant with severe anemia who had human parvovirus B19 infection.
Case Presentation: A male newborn with gestational age of 36 week was born from a mother with poor prenatal care and history of contact with domestic animal. The neonate was very pale with Apgar score 2 at 1 min and received resuscitation, mechanical ventilation and repeated blood transfusion The hemoglobin level was significantly low. Analysis was made based on the clinical presentations. According to the case history, physical and laboratory findings, neonatal severe anemia induced by parvovirus B19 infection was suggested and Laboratory work up documented his infection with parovirus B19.
Conclusion: Parvovirus B19 (B19 virus) is the smallest single strand linear DNA virus in animal viruses, which is the only strain of parvovirus that is pathogenic in humans. Human parvovirus B19 may cross the placenta and result in fetal infection, morbidity and death. Parvovirus is an uncommon cause of neonatal anemia and hydrops fetalis so this etiology must be considered in differential diagnosis of anemia at birth.
Jila Yavarian , Nazanin Zahra Shafiei Jandaghi, Farhad Rezeai , Talat Mokhtari Azad,
Volume 72, Issue 1 (4-2014)
Abstract
Background: Influenza viruses are one of the most important etiological agents of res-piratory disease in humans and cause epidemics and pandemics with substantial mor-bidity and mortality worldwide. Vaccination and antiviral treatments are the sole and essential way for the prevention and control of influenza infection. During an influenza epidemic before the production of effective vaccine, antiviral treatments are the first step for the prevention and treatment of influenza infection. Adamantanes and neuraminidase inhibitors are influenza antiviral drugs. Because of the increase of drug resistant viruses, the aim of this study was the evaluation of the antiviral drug resistance in influenza A/H3N2 viruses from 2005-2013 in Iran.
Methods: In this study 50 influenza A/H3N2 viruses isolated in cell culture were tested. All samples were subjected to M and NA gene sequencing at the National Influenza Center, School of Public Health, Tehran University of Medical Sciences. RNA was ex-tracted from 200 µl of cell culture supernatants using the Roche high pure viral nucleic acid kit. RT-PCR with the Qiagen one step RT-PCR kit was done. The expected size of the PCR products were analyzed by electrophoresis using 1% agarose gels. The PCR products were sequenced for finding the drug resistant mutants.
Results: All influenza A/H3N2 viruses except four viruses circulating during 2005-2006 had Ser31Asn mutation at M2 channel protein. In the analysis of neuraminidase gene none of the A/H3N2 viruses had K292R, E119V and N294S mutations responsible for drug resistant strains.
Conclusion: This study showed circulating A/H3N2 viruses was resistant to adaman-tanes but susceptible to neuraminidase inhibitors. The national data analyzed in this re-search may help increase knowledge about influenza virus antiviral drug resistance, which is a global public health concern. The authors suggested continuing this study and also the investigation of antiviral drug resistance of influenza A/H1N1 and B viruses.
Khadijeh Fanayi, Mehdi Ajorloo , Sayed Hamid Reza Mozhgani , Shiva Irani , Alireza Gholami ,
Volume 72, Issue 5 (8-2014)
Abstract
Background: Rabies is an acute encephalitis that causes more than 60,000 deaths worldwide. The only way to save individuals bitten by a rabies-infected animal is the timely use of effective vaccines. Treatment with new generation vaccines is expensive. Therefore, there is a global movement towards the production of less expensive vaccines which retain and improve upon the quality and effectiveness of the vaccine. Production and evaluation of non-classical vaccines is one of the approaches taken in this regard. In this study, we describe a new eukaryotic expression system to express the nucleoprotein N gene of rabies virus which, if suitable, may be evaluated for anti-rabies vaccine production.
Methods: The complete sequence of the N gene of rabies virus PV subtype was amplified by real-time polymerase chain reaction and cloned into the pCDNA3.1(+) vector. The cloned gene was excised from the vector by restriction enzyme digestion and sequenced. Due to mutations detected in the N gene, the gene coding sequence was purchased as a recombinant pGH/N vector. Vector pGH/N was amplified and following enzymatic digestion, the excised N gene was once again cloned into vector pCDNA3.1(+). Successful cloning was confirmed using restriction digests and quick check. The recombinant vector pCDNA3.1(+)/N was transformed into cultured BSR cells and protein N expression was analyzed using fluorescent antibody test (FAT).
Results: Electrophoresis confirmed amplification of the nucleoprotein N gene and subsequent restriction enzyme digestion showed that the N gene had been successfully cloned into the recombinant pCDNA3.1(+)/N vector. However, DNA sequencing revealed the presence of mutations within the N gene. Restriction digest of the commercial pGH/N vector showed that the N gene had been excised from the vector. Successful cloning of the N gene into the pCDNA3.1(+) expression vector was confirmed using restriction digests and quick check. Protein expression in BSR cells was assayed by immunostaining with anti-ribonucleocapsid FITC-conjugated antibody and visually confirmed by fluorescence microscopy.
Conclusion: This study showed that the protein N of rabies virus subtype PV can be expressed in a eukaryotic expression system using the pCDNA3.1(+) expression vector.
Sina Soleimani , Shahla Shahsavandi , Omid Maddadgar , Homayoon Mahravani , Mohsen Lotfi ,
Volume 73, Issue 3 (6-2015)
Abstract
Background: In the last decade due to emerge and remerge of influenza viruses, quality improvement of vaccines to increase immune responses in target populations have been more necessary. The potential of biologic adjuvant to stimulate and induce immune system is the basis of modern researches in prevention and controlling program of infectious diseases. In this study, the effect of the coding sequence of cellular Myxovirus resistance (Mx) protein as a biological adjuvant for inducing humoral immune response against influenza virus was investigated.
Methods: The experimental study was performed on Balb/c mice in Razi Vaccine and Serum Research Institute from June to November 2014. Three conserved motifs of Mx were selected following sequence alignment between human, mouse and bird species. Potential of the motifs for stimulation immune responses against influenza virus were evaluated using in silico analysis. Based on the immune informatics data Mx1 sequence was the best immune inducer and cloned into pcDNA3.1 vector. Then formulated with inactivated H9N2 influenza antigen as adjuvant and injected to mice groups. The sera of vaccinated mice were collected prior to priming and boosting injections and also at defined weeks and analyzed with serological assays. Histopathological examination was done for evaluation of the vaccine and adjuvant safety.
Results: The mean weight of the Balb/c mice in all control and treatment groups was similar and ranged from 21 to 37 gr (P= 0.05). The difference in increasing antibody titers against influenza virus in immunized mice who received Mx1-adjuvanted vaccine especially in second boosting was significant (P= 0.01) compared to the vaccine alone group. More than 78% of the immunized mice receiving two-time boosting have the mean antibody titer of >6 (Log2) which was higher (P= 0.001) comparing to the mice with one booster injection.
Conclusion: These data revealed that Mx1 as biological adjuvant was able to increase antibody titer and induction memory immune responses against influenza immunization without causing any side effects.
Somayeh Zamani, Fatemeh Fotouhi Chahouki, Zahra Nourmohammadi , Saeideh Sadeghi Neshat, Vahideh Mazaheri , Ali Torabi , Behrokh Farahmand ,
Volume 73, Issue 7 (10-2015)
Abstract
Background: The influenza virus is one of the most important factors for higher morbidity and mortality in the world. Recently, researchers have been focused on influenza conserved antigenic proteins such as hemagglutinin stalk domain (HA2) for vaccine production and serological studies. The HA2 plays a major role in the fusion of the virus with host cells membrane. The immunity system enables to produce antibody against HA2. The aim of this study is polyclonal antibody production against influenza HA2. Methods: This study was done in the Influenza Research Lab, Pasteur Institute of Iran, Tehran for one year from September 2013 to October 2014. In the present study, recombinant HA2 protein was produced in prokaryotic system and purified using Nickel affinity chromatography. The purified HA2 was mixed with Freund’s adjuvant (complete and incomplete) and injected into two New Zealand white rabbits by intramuscularly and subcutaneously routes. Immunization was continued for several months with two weeks interval. Before each immunization, blood was drawn by venous puncture from the rabbit ear. Function of rabbit's sera was evaluated using radial immunodiffusion (RID) in both forms, Single RID (SRID) and Double RID (DRID). Finally, antiserum activity against HA2 was evaluated using western blotting as serological assay. Results: Sedimentary line and zone was observed in RID assays (SRID and DRID) represent interaction between HA2 protein and anti- HA2 antibody. As well as, western blotting results was positive for HA2 protein. Therefore, these results showed that polyclonal antibody produced against HA2 protein can identify HA2 protein antigenic sites. Conclusion: These findings show that humoral immune responses have properly been stimulated in rabbits and these antibodies can identify HA2 protein and may be suitable for other serological methods.
