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Abstract Background and objectives: HTLV-1 virus belongs to the retrovirus and infection with this virus mostly is seen among people having more than one time blood transfusion. Because of requiring repeated blood transfusions, thalassemic patients are considered to be high risk subjects in this regard. Thus, this study was carried out to indicate the frequency of HTLV-1 infection among the thalassemic patients. Materials and Methods: Blood samples of 181 thalassemic patients referred to Taleghani hospital during nearly two years (2004-2005) were taken. By using ELISA technique, the sera were assessed to determine HTLV antibody. The positive ones subsequently were examined by western Blot (kit, 2.4) to confirm the ELISA positive samples and also to recognize the HTLV type. Results: Of 181 thalassemic patients, 93 (51.4%) were male. The age was between one and twenty five (14.11 ± 6.5). 93.4% (169) were received packed cell only once in a month. 14.9% (27) were HTLV positive by ELISA technique, while just eight out of these 27 were considered to be true positive by Western blot and to be contaminated by type one virus. Of all subjects, 4.4% were positive HTLV1. Furthermore, the contamination with this virus is increased as the patients getting older. Conclusion: The findings indicated that among the thalassemic patients in Gorgan, there are cases with HTLV-1 whose frequency is correlated with the other part of our country. Consequently, further comprehensive studies are required to identify those infected blood donated to minimize the transmission risk of this infection in the society and in particular among the people receiving blood, such as thalassemic patients. Keywords: HTLV-1 antibody, thalassemic patients, ELISA, western Blood, Gorgan Journal

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Abstract Background and objectives: Hemin is a porphyrin compound derived from hemoglobin, the precursor of other porphyrin hemoglobin derivatives and the raw material of Hematin. Since hemin is widely used in medicine, we decided primarily to synthesize this substance in Laboratory and to determine the best way of hemin extraction from untransfused and expired blood units. Materials and Methods: In the first method, Glacial acetic acid and sodium chloride were added to citrated blood and hemin crystals were extracted by means of cooling. Finally, the obtained product, by visible spectrophotometer and Infrared Spectrophotometer, was compared to standard samples. Fur thermore, citrated blood, citrated blood hemolysed by distilled water and citrated blood washed by normal saline were used comparatively as a raw material to produce Hemin. The second method was performed by adding Strontium, acetic acid and acetone to blood samples and then after precipitating Hemin crystals they were washed and dried with acetone. Results: The presence of functional groups in Hemin samples, analyzed by infrared Spectrophotometer, indicates the production of this compound. The results of visible Spectrophotometer in comparison with control samples and the results of samples weighting demonstrates high efficiency of extraction stages and the purity of obtained compound. Conclusion: The use of intact citrated blood produces more Hemin than the other kind of Citrated blood samples. Moreover, acetic acid with citrated blood, without any processing on blood, is the best way for Hemin production. Key words: strontium, Hemin, Blood, acetic acid, extraction

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Abstract: Introduction: Influenza is highly transmitted disease and vaccination is the most effective way to prevent influenza. This research was designed to study the variation of serum antibody level among the subjects had already been vaccinated against influenza. Materials and Methods: This research is a descriptive-analytical study, which was carried out on 196 subjects who had influenza vaccination (influvac 2005/2006) and 200 subjects matched by the vaccinated subjects, by age. The subject's serums were prepared seven weeks after influenza vaccination, and the control group's serums were also prepared. The serum antibody level was determined by haemaglutination inhibition test. Results: The mean age of case group is 52.2±11 and control group 48.64±5.17.The antibody titre of 115 of Vaccinated group and 15 of control is less than 40 1 The mean antibody titer of vaccinated subjects and control group is 143.4 ± 10.89 and 18.34± 3.2, respectively. The difference is statistically significant (P value=0.000). Conclusion: The findings show that the mean titer of antibody in vaccinated and control group is statistically different. It means that the influenza vaccine had a good efficacy. Key words: Vaccination, Influenza, Gorgan.

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Abstract False positive results are the major problem influencing interpretation of Clinical Laboratory test. They are originated mostly in the other diseases, technical errors and the recent vaccination. The problem has been presented since 1991 is positive HIV test after influenza Vaccination (1). The False positive has been reported in Cases using Eliza, one of The most common test to screen HIV, and in people vaccinated against influenza.(2) In a study carried out in 1992, nearly 1.7% of recently vaccinated subjects had false positive HIV(3) while Mackenzie's report was 0.6% to 1.7%. (4) In another study, the rate of false positive is 0.9% in 10-20 year-old subjects and 3.1% in subjects aged over 60. (5). In addition, the vaccinated cases suffered from infecfious disorders are highly predisposed to false positive HIV. (5) Considering the aforementioned points, we decided to determine false positive HIV in 196 vaccinated Cases set off for Mecca. (2004, Gorgan). After thorough examination of the cases , we injected them Influac (2005/2006) composing of 45my heamagglutinine and neuramidase proteins extracted from Viruses of: 1-A/California/7/2004 (H3N2)-Like strain (A/New York/55/2004 NYMC X-157) 2-A/New Caledonia/20/99 (H1N1)-Like strain (A/New Caledonia/20/99 IVR-116) 3-B/Shanghai/361/2002-like strain (B/Jiangsu/10/2003) in 196 healthy influenza vaccinated group. Analysis of HIV anti-body was assessed using Elisa method (DiaPro Kit Italy). After seven weeks, HIV anti-body was analyzed using Eliza method (Diapro kit, Italy) Conclusion: The results show that no one has positive HIV. These finding is not in accord with previous studies. It may be due to the recent use of vaccine modified and specialized and the use of Eliza

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Abstract Backgrounds and objectives: Herpes simplex virus type 1 (HSV- 1) infections are mostly shown as a Herpes disease, but It causes conjunctivitis, genital herpes, encephalitis and newborn herpes. This study was conducted to determine the sero-epidemiologic prevalence of herpes simplex virus type 1 in cases referred to clinical laboratories of Gorgan, Iran. Material and methods: In this cross sectional study, we did random blood sampling on 406 cases referred to the Gorgan city's clinical laboratories. These samples were analyzed for HSV-1 Immunoglobulin G and M antibodies using type- specific enzymelinked Immunoassays (ELISA). Results: Of 406 participants, the HSV-1 seroprevalence is 49% (44.3% and 4.7% for IgG and IgM respectively). There is not significant relationship between seropositive HSV-1 and gender, ethnicity, age and marital status. Conclusion: Sero-epidemiological of HSV-1 in Gorgan is the same of the other places in Iran, but it is higher than European and lower than African countries. It seems that the people’s culture is very important. Therefore it needs to be investigated more. Keywords: HSV-1, Antibody, Gorgan.

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Abstract Background and objectives: Human T-Lymphocyte Virus-1 (HTLV- 1) is known as the etiologic factor of acute T-Lymphocytic Leukemia (ATL) and tropical spastic paralysis. (TSP). Endemic factors causing infection with Human T Lymphocyte Virus-1 (HTLV-1) is based on environmental, socio-economical and health behaviors of the individuals. This virus is well distributed in families with involved members. Golestan province is located in North West part of Northern Khorasan province that had already been known as an endemic area for HTLV-1. This virus is also known as the main etiologic factor for cancers and ATL, therefore we studied the prevalence of HTLV-1 seroepidemiology in Golestan province. Material and Methods: The subjects selected by cluster sampling were 2034 healthy cases residing in different parts of Golestan province. ELISA method using Dia- pro anti HTLV-1 antibody kits was applied for serological assessment. Western Blot (HTLV BLOT 2.4) was used for confirmation purposes. Results: The subjects aged 38.66±16.54 were 2034 healthy persons. Forty-one point seven of these cases were males and the rest females. Based on ELISA method there were15 HTLV-1 positive cases (0.7%). -1. (0.7%) Six out of 15 were confirmed by western blot method (95%, CI: 0.06-0.53%). The highest prevalence sigllificant) aiology is in the highat rate in 31-40 year old gro0.7%). onclusion: This study shows that HTLV-1 is prevalent in Golestan the same as the other parts of the world. There fere: we urse on performing screening test (HTLV-) on donated blood components before delivering (OK labeling). Key words: HTLV-1, Seroepidemiology, ELISA, Western Blot, Golestan ATL(Acute T lymphocytic Leukemia) Six cases out of 15 were confirmed by western blot method (95%, CI: 0.06-0.53%). The highest prevalence was 2.6% seen in Kalaleh city (east part of the province) [95%, Cl: 0.06-0.53%). There was significant difference between the prevalence of HTLV-1 and the dwelling place. (p=002). HTLV-1 seroepidemiology was in the highest rate in 31-40 year old group (0.7%). Conclusion: This study shows that HTLV-1 is prevalent in Golestan province, the same as the other parts of the world. Therefore, we recommend performing screening test (HTLV-) on donated blood components before delivering (OK labeling). Key words: HTLV-1, Seroepidemiology, ELISA, Western Blot, Golestan province, ATL (Acute T lymphocytic Leukemia)

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Abstract Background and Objectives: Diarrhea is one of the most common diseases in the world. Campylobacter jejuni is one of the prevalent agents of bacterial diarrhea in most of developing countries. It is usually ignored in routine laboratory test in our country, because it has a difficult investigation method. This article aims to determine the prevalence of Campylobacter jejuni, in diarrhea samples in Gorgan City (East north of Iran) by PCR Method. Material and Methods: This descriptive study was performed on 455 diarrheal samples during one year (2005-06).255 out of them were cultured on Preston media (Himedia co.) on 42°c. DNA Extracted by phenol cholorophorm method was directly carried out on stool samples.16srDNA hipo and asp primers for detection of Campylobacter genus, C.jejuni and C.coli species were used, respectively. In addition, universal primer of 16srDNA was used for control of PCR method. Results: no sample was positive for Campylobacter in culture .only three samples were positive for Campylobacter genus and C.jejuni specific primer but none of them were positive for C.coli .99 samples were positive by universal primer of 16srDNA . Conclusion: The results indicate that C.jejuni isn't a prevalent agent in diarrhea in our region. Key words: Campylobacter jejuni -Gorgan- Diarrhea

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Abstract Background and objectives: Infection has a Leading role in pregnancy. Cytomegalovirus (CMV), listeria and Toxoplasma are the most common causes of infection in human. Based on the previous researches, about 15-25 percent of being infected during pregnancy leads to some complications such as abortion, fetal death, early labor and etc. This study was designed to determine the seroprevalence of Cytomegalovirus (CMV), Toxoplasma gondii and Listeria moncytogenes among pregnant women in Gorgan, north of Iran (2005-2006). Material and Methods: we conducted this Simple randomized study on 118 unsuccessful pregnant woman and 99 successful ones referred to Deziani hospital in Gorgan. We assayed both IgG and IgM antibodies for CMV and Toxo by Elisa and IFA method for Listeria. In addition, we fill out a Check list and then use SPSS soft ware, chi square to analyze the data. Results: The frequency of IgG for CMV and Toxo is 89.9% and 45.5% in successful pregnant women and 77.1% and 44.1% for unsuccessful pregnant women (P=0.41, P=0.01). IgM frequency for CMV and Toxo is 14.1% and 46.5% in successful women and 30.5 and 21.7% in unsuccessful ones. (P=0.003, P=0.002)Total frequency (IgG, IgM) for Listeria is 7.62% and %3.03 in successful and unsuccessful women, respectively. There is a significant relation between abortion and IgM titer against Toxoplasma in successful and unsuccessful groups. (P=0.003).This relation is true for total antibody titer against Listeria (P=0.003). Conclusion: Because of high titer of antibodies against CMV, Toxo and Listeria in unsuccessful pregnant women, suffering from these agents during pregnancy may result in abortion and fetal death. Hence, we recommend to hold some preventive and educational program and also to assay antibodies against theses agents. Key words: Listeria moncytogenes, Cytomegalovirus (CMV), Toxoplasma gondii, success and non-success pregnancy, Serology, Gorgan

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Abstract Background and objectives: Breast cancer is the most prevalent one in women. Some of the common causative factors are genetic background, nutritional and environmental factors. Viruses are believed as a risk factor in this cancer, too. Recent studies reported that Human Papillomaviruses can be one of the possible risk factors of breast cancer. This study focused on investigation of the papillomavirus genome in tissues of breast cancer in Golestan province, Iran. Material and Methods: This descriptive analytical study was done from 2005 until 2008. The Samples were obtained from women admitted to the hospitals in Gorgan and Gonbad cities. All breast biopsy or mastectomy tissues were confirmed by the pathologists for breast cancer. DNA was extracted and PCR done by using general primers (GP5 + / GP6 + and MY09/MY11) for detection of papillomavirus genomes. Results: The Subjects are 231 patients aged 47± 12/72, the youngest 20 and the oldest 84. They are from Gorgan (N=122)and Gonbad (N=109) The result Shows That The Subjects Suffer from infiltrating ductular Carcinoma(31.4%), infiltrating duct Carcinoma (60.1%)and intraductal Lobular Carcinoma (4.3%) and The rest from other kinds of Cancer. Papilloma Virus genome is not found in These Samples. Conclusion: Based on paradoxical results from different parts of the world, upon the presence or absence of papillomavirus genome in breast cancer samples, to show the role of this virus in the development of breast cancer more studies are needed. Key words: Breast Cancer, Papillomaviruses, Golestan Province

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Abstract Background and objectives: With almost nine million new cases each year, tuberculosis is still one of the most Life-threatening diseases in the World. Distribution of drug resistant strains of M.tuberculosis has a lot of importance. This research was carried out to determine the frequency of drug resistance of M. tuberculosis in strains isolated in Golestan province. Material and Methods: In this cross -sectional study, 104 isolate of M.tuberculosis which isolated from patients referred to Gorgan tuberculosis Health Center, in 2008 were studied. DNA was extracted by Boiling Method. By using PCR method, we determine the M.tubeculosis strain and resistance to Rifampin (Using IS6110 and Gene rpoB primers) and resistance to Isoniazid (Using InhA and KatG primers). As a Gold Standandard, “Proportional method” was performed for 45 Samples. Results: 87 strains were identified as M.tuberculosis. 6.9% of them were resistant to Isoniazid, 4.6% to Rifampin and 2.3% to both (MDR).Sensitivity and Specifity of PCR method in detection of resistant to Isoniazid were 95.3% and 57.1% and for Rifampin were 94.7% and 33.3%. Conclusion: We found that in our region, the MDR is not very common. More than 16% of isolated strains from tuberculosis suspected patients were MOTT, for this reason it is necessary to mention that use biochemical or PCR method to determine M.tuberculosis is necessary. Key words: Mycobacterium tuberculosis, MDR, PCR, Proportional method , Golestan province.

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Background and Objectives: rapid, accurate and cost effective diagnosis of infectious and non infectious diseases is an essential step for treatment process. Nowadays, in Line with scientific progression in molecular biology, genetics and biochemistry which are based on biotechnology and genetic engineering aspects, new branch of medicine entitled molecular medicine is being derived. It can be helpful in three areas of diagnosis, prophylaxis and treatment. This new branch is going to identify further complexity of diseases and to present efficient solutions for growing health criteria. Therefore, updating and being familiar with the new procedures related to diagnosis, prophylaxis and therapy are necessary for our society. In this paper, we are trying to introduce NASBA technology which has a high potential, at genome level, in recognizing specific characteristic and unique genetic markers of microorganisms. This technology has numerous benefits for easy detection of infectious diseases such as tuberculosis. Furthermore, we review the methods of tuberculosis detection.

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Abstract Background and objectives: Mumps virus is one of the first known causative agents of meningitis in children. On-time diagnosis is the first step in treating meningitis. We aimed to evaluate Mumps virus meningitis in children in Gorgan, Iran Material and Methods: CSF and blood samples were taken from children with meningitis, Jun 2008 till Sep 2010. For 40 samples with negative bacterial culture, Extraction of viral RNA was carried out and Real-time PCR was performed for detection of Mumps virus. Demographic, clinical, biochemical and cytological data were collected. We run SPSS version 18 to analyze the data, using Chi Square (p<0.05). Results: three (7.5 %) samples have Mumps virus, two boys and one girl. All three positive cases have 0.5-1 degrees Celsius fever and vomiting but no bulging fontanel. They have not Kernig, Rodor, Brudzinski’s sign, hepatosplenomegaly, lymphadenopathy, pharyngitis and rash. ESR is higher than normal in all positive cases and CRP is positive in two cases. Protein of CSF in one case is higher than normal range. Conclusion: meningitis is an emergency condition therefore, molecular diagnostic techniques are recommended for early diagnosis and intervention. Key words: meningitis, mumps virus, cerebrospinal fluid, Real-Time PCR

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Abstract Background and objectives: the increasing use of antibiotics, especially the third generation cephalosporins, is an important factor in the spread of antibiotic resistance in bacteria. The main reason for the development of resistance phenotype such as Extended Spectrum Beta Lactamas (ESBL) is the extensive use of broad-spectrum cephalosporins. In phenotypic survey, the Phenotyping confirmatory test and the minimum inhibitory concentration (MIC) are used. In this study, the prevalence of the isolates resistant to third generation cephalosporin (cefotaxime) was determined based on MIC. Material and Methods: form September 2010 to September 2011, 75 isolates of Klebsiella pneumoniae were collected from the infections of inpatients and outpatients, referred to state and private laboratories of Gorgan. For all of the Klebsiella pneumoniae strains, MIC determination using E-test (company Liofilcheme-Italy) was performed. Results: According to the MIC results, 26 samples (34.6%) are resistant to cefotaxime 22 isolates are completely resistant to concentration of 256μg. Conclusion: Because of the importance of risk of becoming ESBL, further studies are needed to clarify the ESBL in the region. Keywords: ESBL, MIC, Klebsiella pneumoniae, Cephalosporin

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Abstract Background & Objective: Hepatitis E virus is one of the most common causes of acute infection in adults. Pregnant and transplant patients are more in risk of HEV infection. Fecal-oral is the main route of HEV transmission but recently transmission by blood transfusion has been observed. This study was aimed to determine the prevalence of HEV-Ab in hemodialysis patients in Gorgan, Iran. Material and Methods: In this cross-sectional descriptive study, we investigated 150 hemodialysis patients of Panje Azar hospital in Gorgan. These patients were evaluated for the presence of HEV total Ab by ELISA method. Results: of 150, 6 patients (4%) are positive for HEV-Ab. There has been no significant relation between anti HEV Ab and variables such as age, gender, ethnicity, duration and number of hemodialysis in a week and (P>0.05). Conclusion: This study, which is the first report from this area, show that the lower prevalence of anti HEV Ab in hemodialysis patients in comparison with pregnant and childbearing age women. Keywords: Hepatitis E Hemodialysis Elisa Gorgan

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Abstract Background and Objective: Resistance to antiretroviral agents is a significant concern in clinical management of HIV-infected individuals. Resistance is the result of mutations that develops in the viral protein targeted by antiretroviral agents. Material and Methods: In this cross-sectional study, the blood samples of 40 HIV-positive patients were collected. Twenty of them were drug-naïve and the rest were under treatment for at least one year by antiretroviral agents. Virus genome was extracted from patient's plasma with high-pure-viral-nucleic-acid kit. Then, by means of reverse-transcriptase and specific primers of protease genes were amplified and sequenced. Sequences of genes, drug- antiretroviral- resistant mutations and subtypes were determined using Stanford University’s HIV-drug-resistance databases. Results: Drug-naive patients show 15% resistance to nucleoside-reverse-transcriptase inhibitor (NRTI) and 20% resistance to non-nucleoside-reverse-transcriptase inhibitor (NNRTI). Anti-protease resistance is not observed in any patients. In under treatment patients, drug resistance to NNRTI (25%) is more than drug resistance to NRTI (20%) and the rate of drug resistance to protease inhibitor is 5%. Conclusion: Our findings show a high prevalence of drug-resistant mutations in Iranian-drug-naïve-HIV-infected patients. But in under treatment individuals, the rate of drug resistance is less than previous studies. Keywords: HIV Nucleoside Inhibitor Non-Nucleoside Inhibitor Protease Inhibitor

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Abstract Background and Objective: The emergence of a novel H1N1influenza A virus of animal origin with transmissibility from human to human poses pandemic concern. Current subtypes of Seasonal influenza A viruses spread in human are influenza A H1N1 influenza A H3N2 and influenza type B viruses. The aim of this study was to determine current strains of the H3N2 and new H1N1 subtypes of influenza A virus from patients suspected influenza infection in 2009 flu pandemic in Golestan province, Iran. Material and Methods: In this descriptive study, respiratory samples (n = 153) from patients with acute respiratory symptoms were collected in 2009 flu pandemic applied during 2009 pandemic influenza in Golestan province. After reverse transcription of extracted viral RNA, PCR was developed for both H1N1and H3N2subtypes using CDC specific primers. Results: The mean age of patients was 16.59. Of them 45.1% were male. Thirteen (8.49%) were infected with seasonal influenza H1N1 and 25(16.33%) with seasonal H3N2influenza. Conclusion: The rate of infection with seasonal H1N1and H3N2is similar to other studies reported from Iran, but lower than the rate reported from other parts of the world. Key Words: Influenza A Virus, H1N1, H3N2, RT-PCR, Iran

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Background and Objective: Cytomegalovirus (CMV), one of the most common opportunistic pathogens in patients infected with human immunodeficiency virus (HIV), can cause the diseases such as encephalitis, pneumonia, and chorioretinitis. This study aimed at molecular studying of CMV infection in individuals infected with the human immunodeficiency virus. Material and Methods: In this study, 50 venous blood samples from HIV-infected individuals were taken. Patients were divided into two categories: patients under treatment with and without antiretroviral drugs. Plasma were separated from blood samples and examined for the presence of cytomegalovirus genome by PCR. Material and Methods: this study was conducted on 50 blood samples from HIV-infected individuals, and plasma was separated and examined for the presence of cytomegalovirus genome by PCR. Patients were divided into two group of under treatment with and without antiretroviral drugs. Results: Of 50, 28 (% 56) were men and 22 (% 44) were women. CMV genome was identified in 8 samples (16%), and the molecular prevalence of CMV infection was 21.4% (n= 6) in males and 9.1% (n = 2) in females. Conclusion: Given the frequency of Cytomegalovirus Active Infection in HIV-infected individuals under antiretroviral therapy, we should be careful about the treatment of Cytomegalovirus Active Infection. Keywords: Active Infection, Cytomegalovirus, Human Immunodeficiency Virus, Shiraz, PCR

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Abstract Bachground and Objective: most of environmental microorganisms have the genes resistance to antibiotics and metals. The aim of the current study was to survey resistance pattern to some antibiotics and heavy metals in three pseudomonas aeruginosa isolated from different ecological areas. Material and Methods: first, the isolates were identified by biochemical methods and phylogenetic analysis. Then, the evaluation of antibiotic resistance was conducted by disc diffusion and that of Heavy metal resistant by agar dilution, in a range of 50-500 µg/ml. Results: The results showed that all three isolates were resistant to beta lactam antibiotics. Although these isolates were highly resistant to heavy metals, no relationship was observed between ecological sources and the resistance pattern in ICT1 and Abt2 strains. However, strain Q isolated from digestive system of ParmacellaIberica showed high resistance to antibiotics and low resistance to heavy metals. Conclusion: given that environmental bacteria have a high potentiality for carrying resistance genes and this can be an advantage environmentally, they could be used to remove heavy metals from polluted areas. On the other hand, resistance genes medically are a concern due to probability of transferring to pathogen strains. Keywords: Antibiotic Resistance, Heavy Metal Resistance, Pseudomonas Aeruginosa

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Abstract Background and Objective: Lamivudine is the first orally available drug approved for treatment of chronic hepatitis B. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene contribute resistance to lamivudine. This study was aimed to determine the rate of YMDD and FLLAQ mutants in hepatitis B patients in Golestan Province, Iran. Material and methods: In this cross sectional study, 120 patients with chronic HBV infection were recruited. Of them, 55 were treated and 65 untreated with Lamivudine. HBV DNA extractions from plasma and polymerase chain reaction (PCR) were performed. For detection of Lamivudine mutants direct sequencing and alignment of products were applied using reference sequence from Gene Bank database. Results: the average age of patients was 36.31±10.07, which 35% of them were female and 65% were male. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene were detected in 12 of 55 patients (21.81%) treated with Lamivudine while no mutation was observed in in untreated patients. The YMDD and FLLAQ mutants were detected in 9.16% (11/120) and 0.83% (1/120) of chronic HBV patients, respectively. Conclusion: Usual HBV mutations, which play an important role in lamivudine resistance, detected in this study are similar to other studies. Key words: Hepatitis B Viruse, YMDD Mutation, Lamivudine, Iran.

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Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative