Author: Ruhollah Dorostkar

Introducing two effective techniques for the detection of adenovirus respiratory infection in human sample

Recently, respiratory viral infections have witnessed substantial diagnostic evolution, with the advent of emergent pathogenic agents and the improvement of new diagnostic approaches. Acute viral respiratory infection (ARVI) commonly causes illness and fatality, particularly in pediatric patients. Adenovirus has the highest prevalence in upper and lower respiratory tract infections (RTIs) amongst respiratory pathogens of importance. The purpose of this research was to introduce two effective techniques for the detection of adenovirus respiratory infection in Human samples, cell culture, and real-time PCR. Samples from patients aged less than ten years were obtained from Baqiyatallah (as) Hospital and cultured on HEK cells after preparation. After observing the cytopathic effects of the virus, a molecular test (Real-time PCR) with the help of specific primers was done on samples taken from the cell culture. Both assays detected viruses in clinical samples, and results of cell culture and real-time PCR confirmed infection with adenovirus in samples. According to our findings, simultaneous use of tests (cell culture and PCR) based test real-time PCR is a somewhat reliable method for detecting adenovirus in clinical specimens.

Gene therapy by using stem cells in the treatment of hepatitis

Chronic hepatitis can resolve spontaneously or progress to more advanced stages, i.e. fibrosis, cirrhosis, and liver cancer, respectively. Two million deaths each year are attributed to liver diseases globally. Chronic liver disease and cirrhosis comprise 11th position in leading causes of death worldwide. Viral infections account for the etiologic factor of most cases of hepatitis. Almost the only available way for the treatment of end stages of hepatitis is liver transplantation. The biggest problem with this therapeutic approach is the number of liver transplant donors is less than the number of recipients. Therefore, more advanced methods must investigate. Recently, gene therapy by the utilization of stem cells yielded promising results in clinical and preclinical investigations to the treatment of liver diseases. Novel genome editing tools such as TALENs and CRISPR/Cas9 consider as powerful genome correction approaches and can employe for iPSCs gene editing. This paper highlights the various aspects of gene therapy through stem cells for liver diseases

Development of quantitative real-time RT-PCR assay for detection and viral load determination of Crimean-Congo Hemorrhagic Fever (CCHF) virus

Background and Aim: The CCHF (Crimean-Congo hemorrhagic fever) virus causes a severe disease in human with a case fatality rate of up to 50%. Since, there is no specific treatment or approved vaccine against CCHF viral infections, an accurate and early detection as well as a reliable surveillance and quantitative determination of viral load is necessary for patient improvement and case management. In this research, our aim was to develop a probe based one-step real- time reverse-transcription polymerase chain reaction (rRT-PCR) assay for in-house quantitative detection of CCHF virus. Methods: At first, the highly conserved S-fragment sequence of CCHF virus genome was adapted from GenBank and the specific probe and primers targeting this region were designed. Then, viral RNAs were extracted from 37 blood samples of different patients from east of Iran (Zahedan). The specificity and sensitivity of the probe and primers were also evaluated in positive blood samples, confirmed to have CCHF virus. A standard (PTG19-T vector containing S-fragment) for quantization was also constructed and the viral load was determined in some of positive samples. Results: From a total of 37 suspicious blood samples, 15 samples were confirmed to be positive for CCHF virus by this probe based one-step rRT-PCR assay and no false-positive result was detected according to sequencing data. The predicted fragment of 176 bp was also confirmed in all positive samples by gel-based electrophoresis analysis. The assay was linear between 10 to 103 copy numbers per each microliter of extracted plasmid for this technique and the viral load determined in one of patient blood samples was 55,000 viral particles per each milliliter, for example. Bioinformatics and experimental evaluations approved the specificity of this assay. The LOD of the assay was 10 (or fewer) copy numbers of viral genome per each microliter of the extracted genome. Conclusions: This research showed that the developed probe based one-step rRT-PCR assay is a specific, rapid, sensitive and the simple tool for detection and viral load determination of the CCHF virus.

Chest CT-scan findings in COVID-19 patients: the relationship between the duration of symptoms and correlation with the oxygen saturation level

Purpose: This study is carried out to evaluate the diagnostic value of using common features of computed tomography (CT) imaging in COVID-19 disease, and to assess the relationship between blood oxygen saturation level and severity of CT findings. Materials and Methods: In this retrospective study, the chest CT of 173 test-confirmed COVID-19 patients have been evaluated to determine the patterns of involvement in multiple phases of illness. Then, the correlation between the severity of lung involvement and oxygen saturation levels has been assessed. Results: The chest CT results show that 87.6% of patients had GGO, which was the most common pattern in our findings. 83.8% of patients had bilateral lung involvement with the dominant multifocal and peripheral distribution. peribronchovascular involvement was also a common finding in our study (47.2 %). we found predominantly peribronchovascular view in 3 patients (1.7%), pleural effusion in 4 patients (2.3%), lymphadenopathy in 10 patients (5.8%), the tree in the bud in one patient (0.6%), and nodules in 4 patients (2.3%). We also found that GGO is the most common pattern during the early phase of the disease (97.4% of early phase cases). However, in the intermediate and late phases, consolidation and crazy paving patterns are more common. Moreover, our findings indicate that there is a significant relationship between oxygen saturation level and Total Severity Score, with the exclusion of the young adult patients (20-40 years). Conclusion: Relying on chest CT-scan findings apart from the oxygen saturation level is sufficient for the diagnosis and management of patients with COVID-19 pneumonia.