Cloning of the Core Antigen (HBcAg) Gene of Hepatitis B Virus into an Eucaryotic Expression Vector

In the presence of this study, the core antigen (HBcAg) of hepatitis B virus (HBV) was amplified and cloned into an eucaryotic expression vector. For this aim, HBcAg DNA was amplified with the primers spesific to HBcAg gene of HBV by polymerase chain reaction (PCR). Afterwards, the PCR product of HBcAg gene of HBV was placed into pcDNA 3.1/V5 His-TOPO vector. PCR screening assay was performed to confirm the presence of the HbcAg gene. Furthermore, enzyme digestion assay was performed for the same purpose. In conclusion, the HBcAg gene of HBV was cloned into the eucaryotic expression vector

Farklı Örneklerden Elde Edilen DNA Parmak İzi Çalışması

Son yıllarda adli tıp alanında, elde edilen materyalin kimliklendirilmesinde, DNA parmak izi büyük bir potansiyel göstermiştir. Bu çalışma: DNA parmak izi tekniğinde farklı biyolojik örneklerin kullanılması ve sonuçların yorumlanması amacıyla yapılmıştır. Bunun için, akraba olmayan 3 kişinin farklı örnekleri (kan, kıl kökü ve tükrük) ile bir kişinin bunlara ek olarak tırnak ve semeninden DNA izole edilmeye çalışılmış, en çok DNA (4.46 pg) semenden, en az da tükürük (0.82 pg) ve tırnaktan (0.90 pg) elde edilmiştir. DNA’lar genomda sıra halindeki tekrarlan (Tandem Repeat) ortaya çıkarabilen bir primer varlığında Polimeraz Zincir Reaksiyonu (PCR) ile çoğaltılmış, %1.4’lük agaroz jel elektrofo- rezinde yürütülmüştür. Böylece aynı kişinin farklı örneklerinde aynı, farklı kişilerde de ayrı motifte DNA ürünlerinin ortaya çıktığı gösterilmiştir. Sonuçta, Elazığ’da da benzeri çalışmaların yapılabileceği ve günlük kullanıma sunulabileceği görülmüştür. Anahtar Kelimeler: DNA parmak izi, DNA izolasyonu, adli tıp

Detection of Molluscum contagiosum virus (MCV) subtype I as a single dominant virus subtype in molluscum lesions from a Turkish population

Background. Molluscum contagiosum has a worldwide occurrence and its primary mode of transmission is via direct human contact including sexual means. The aim of the study was to implement a polymerase chain reaction-based assay for detection and subtyping of Molluscum contagiosum virus (MCV) in skin lesions diagnosed with molluscum contagiosum in a large regional teaching hospital in Turkey

Prevalence and genotypes of hepatitis G virus among hemodialysis patients in Eastern Anatolia, Turkey

Objectives: To study the prevalence and genotype distribution of hepatitis G virus (HGV) in hemodialysis patients in East Anatolia, Turkey. Subjects and Materials: Eighty-nine hemodialysis patients and 30 healthy individuals were analyzed by using reverse-transcriptase nested polymerase chain reaction with primers specific for 5' untranslated region. HGV genotyping was performed by PCR and three randomly selected HGV-positive samples were sequenced. Results: Of the 89 hemodialysis patients, HGV RNA was detected in 9 (10.2%). All of our isolates were assigned to genotype 2. Conclusion: Our results showed that hemodialysis patients carry the risk for HGV infection in East Anatolia, Turkey. Copyright (C) 2005 S. Karger AG, Basel

Investigation of Epstein-Barr virus DNA in formalin-fixed and paraffin-embedded breast cancer tissues

Objective: To investigate etiological role of Epstein-Barr virus (EBV) DNA in breast cancer. Materials and Methods: The presence of EBV DNA in 57 breast cancer tissues was investigated with a sensitive PCR assay. The breast cancer tissues were from invasive ductular (n = 28), lobular (n = 20) and other miscellaneous carcinomas (n = 9). Tissues from normal breasts and patients with various benign breast diseases (n = 55): fibrocystic disease (n = 34), fibroadenoma (n = 16), hyperplasia, and granulomatous mastitis (n = 5), were used as control samples. Results: EBV DNA was detected in 13 (23%) cancerous tissues (7 ductular, 4 lobular, 2 other carcinoma) and 19 (35%) in the control tissues. The difference between EBV presence in malignant and benign tissues was not statistically significant (p > 0.05). Conclusion: The presence of EBV DNA was detected almost equally in both breast cancer and normal tissues, which indicates no etiological role for EBV in breast cancer. We suggest further etiological studies. Copyright (C) 2005 S. Karger AG, Basel

Development of a Novel Plasmid-based Eukaryotic Model to Investigate Crimean-Congo Hemorrhagic Fever Virus

Objective: Crimean-Congo Hemorrhagic Fever (CCHF) is a severe tick-borne viral disease, caused by the Crimean-Congo Hemorrhagic Fever virus (CCHFV). The global expansion of CCHF and high mortality rates underline the critical need for research and development of effective treatments and vaccines. However, the high risk of transmission and requirement for highcontainment facilities hinder investigations involving live virus. In this study, we aimed to address these challenges by employing a plasmid-based virus-like particle (VLP) system and a minigenome model to investigate the biology and immunology of CCHFV. Methods: The plasmids encoding CCHFV structural genes of CCHFV were transfected into Huh-7 cells. Viral protein expression was confirmed using fluorescence imaging, immunological and molecular methods. A minigenome system was developed, eliminating the need for T7 polymerase, T7-expressing cellular lines, or viral ribonuclear protein complexes, allowing autonomous virus replication without a helper virus or transfections using plasmids in trans. Results: Fluorescence microscopy showed successful production of NP-EGFP and GC-EGFP proteins with various subcellular localizations. Western blot analysis demonstrated the presence of pre-Gc, Gc, pre-Gn, Gn, and Np proteins in cell lysates and supernatants. ELISA analysis suggested that transfection of Np alone, in combination with Gc, or all three proteins might cause distinct VLP formations. Huh-7 cells successfully expressed reporter genes after transfection of minigenome RNA transcripts. Conclusion: The study advances CCHFV research by using novel tools for virus biology and immunology. The findings may provide new avenues for research that promise better public health preparation against this neglected viral disease

SV40 in human thyroid nodules

Background: Simian virus 40 (SV40) has been a model experimental system for the study of cell transformation and tumorigenesis for many years. The study of SV40 in humans has aroused interest in the related BK virus (BKV) and JC virus (JCV) and their role in human disease. Objectives: SV40 has been found in a variety of human samples, both malignant and normal. Many independent studies have suggested that SV40 plays a role for some cancers. However, in most cases the role of SV40 remains unclear. Study design: The subject of this study consisted of 99 patients with thyroid nodules. Both thyroid nodule and normal thyroid tissue were taken from each patient to test whether they contained SV40 sequences. Results: We detected SV40 sequences by polymerase chain reaction (PCR) in four of 99 thyroid nodules. Two of them were papillary thyroid carcinomas and the others were benign thyroid nodules. No SV40 was detected in 99 of normal tyhroid tissues of the same patients. DNA sequence analysis, performed in four positive samples, confirmed that PCR products belong to the SV40 T antigen (Tag) region. Conclusion: The possible role of SV40 in the development of thyroid nodules and the spread of SV40 by horizontal infection in the human population are discussed. (C) 2004 Elsevier B.V. All rights reserved

Cross-Reactive Anti-Nucleocapsid Protein Immunity against Crimean-Congo Hemorrhagic Fever Virus and Hazara Virus in Multiple Species

The World Health Organization estimates that there may be three billion people at risk of infection by Crimean-Congo hemorrhagic fever virus (CCHFV), a highly lethal emerging Orthonairovirus carried by ticks. On the other hand, the closely related Hazara virus (HAZV), a member of the same serogroup, has not been reported as a pathogen for humans. Given the structural and phylogenetic similarities between these two viruses, we evaluated the immunological similarities of the nucleocapsid protein (NP) of these two viruses in multiple species. Strong antigenic similarities were demonstrated in anti-NP humoral immune responses against HAZV and CCHFV in multiple species using convalescent-phase human CCHF sera, rabbit and mouse polyclonal antiserum raised against CCHFV, and mouse polyclonal antiserum against CCHFV-NP in enzyme immunoassays. We also report a convincing cross-reactivity between NPs in Western blots using HAZV-infected cell lysate as antigen and inactivated CCHFV- and CCHFV-NP-immunized mouse sera. These results suggest that NPs of HAZV and CCHFV share significant similarities in humoral responses across species and underline the potential utility of HAZV as a surrogate model for CCHFV