Fanleaf degeneration, a disease caused by Grapevine fanleaf virus (GFLV), is considered as the most destructive grapevine viral disease worldwide causing severe losses in vineyards, annually. GFLV is apparently believed to have originated from ancient Persia and therefore, to restrict it in its further infections, a precise identification along with a monitoring of its distribution pattern is crucial. Distribution and relative rate of infection to GFLV in vineyards and nurseries from some different parts of Iran was studied. A total of 882 cuttings were collected from Karaj, Bavanat, Shiraz, Jahrom, Maymand, Ourmia, Naghadeh and Abhar. Infection was confirmed in 204 samples using such different diagnostic methods as mechanical transmission to test plants, as well as serological and molecular techniques. Results confirmed GFLV infection in some new regions of Iran for the first time, including some nurseries in Karaj and some in Naghadeh city. The most severely infected samples were observed in Bavanat, while 63.7 percent of the samples being infected. No infection was detected in Abhar city. Infection varied from 1.8 to 43.1 percent in the other regions. Results proved widespread distribution of GFLV races to different regions of Iran, confirming the fact that it had been spread from its believed origin through infected cuttings and scions.

During 2007-2008 growing season, a total of 520 sunflower leaf samples with symptoms like vein clearing, cholorotic and necrotic local lesion, deformation and stunting which were suspected of being infected with Tobacco streak virus (TSV) were collected from sunflower fields of Azarbayijan-gharbi, Isfahan, Qom, Markazi, Tehran and Hamedan provinces. Infection with TSV in collected samples were determined serologically by enzyme – linked immunosorbent assay (DAS- ELISA) using a polyclonal antiserum. It was determined that 124 samples were infected with TSV. The percentage of TSV infection in sunflower fields of Azarbayijan-gharbi Hamedan, Tehran, Qom, Isfahan and markazi provinces were 56%, 31%, 26%, 14%, 8.8% and 0% respectively. The virus was propagated on maintenance hosts. TSV induced cholorotic local lesions symptoms on inoculated leaves of Chenopodium quinoa, C. amaranticolor and Vigna unguiculata. The virus caused systemic stunting , deformation and curl on leaves of Datura stramonium, Gomphrena globosa, Helianthus annuus and Vicia faba. Nicotiana benthemiana was symptomless. The molecular weight of coat protein under denaturing condition (SDS-PAGE) was estimated to be 30.9 KD and Western blot confirmed the result of SDS-PAGE. IC- RT- PCR and RT- PCR were performed using a specific primer pair of TSV which had been designed according to a part of coat protein, and a fragment with 717 length was amplified.

Soybean mosaic virus (SMV) which belongs to the virus family Potyviridae, causes a disease in soybean that is present in soybean-growing areas of the world, and is widely distributed in northern Iran. Detection of SMV is very important for disease management. In the present study several serological and molecular (nucleic acid- based) methods of rapid virus detection were compared. Serological studies including DASELISA, DAC-ELISA, TPIA and DIBA were optimized and compared to identify the virus by using a polyclonal antibody. Among the serological methods, TPIA and DIBA are simple and TPIA is rapidly and easily applicable in the field. However, TPIA was found to be preferable. TPIA is time-saving, not requiring conventional sap extraction and also nitrocellulose membranes used for printing can be used in the field and stored for a long time or transported to other laboratory to be processed. RT-PCR and Immunocapture RT-PCR (IC-RT-PCR) were performed as molecular methods for detecting SMV using a pair of primers designed to amplify a fragment in the coding region of the SMV coat protein. To extract total RNA for RT-PCR, two methods including RNAWIZ and phenolchloroform were used. A part of the coat protein genome of SMV was converted to cDNA using a reverse transcription (RT) reaction. For IC-RT-PCR method, virus partial purification was carried out by solid-phase (0.2 ml microfuge tube) adsorbed polyclonal antibody, and then the RT reaction was carried out in the tube. In both methods cDNAs were amplified by PCR. Both methods amplified the expected fragment in virus-infected plants. Whereas RT-PCR requires total RNA extraction, ICRT- PCR does not have total RNA extraction problems. Our findings suggest that TPIA and IC- RT- PCR can be routinely used for SMV detection, with high efficiency.

2Background and Aim:  Rapid antigen and antibody, serological tests, and RT ‑ PCR-based molecular methods are widely used to detect microorganisms worldwide. This study aimed to detect the covid-19 using Isothermal nucleic acid amplification techniques. Materials and Methods:  In this study, we collected 200 samples of nasopharynx and oropharynx from Jamaran Heart Hospital in Tehran. Covid-19 was examined by LAMP-RT PCR and Real-Time RT-PCR techniques. The nasopharynx and oropharynx nucleic acids were extracted both by pishtaz RNA extraction kit. LAMP-RT-PCR was performed on direct samples, while Real-Time RT-PCR samples were tested only using RNA extraction samples. The probe-primer mixture of this kit was designed using a dual-target gene method that simultaneously targets the genomic sequences of the spike region and N nucleocapsid. Fluorescence was measured using Real-Time RT-PCR and LAMP-RT PCR. Results: Clinical specimens (56%) were positive using Real-Time RT-PCR technique, with mean Ct values less than 30. Clinical samples (52%) were positive for Covid-19 in the RT-LAMP technique of RNA extraction samples. RT-LAMP technique indicated a sensitivity of 92. 8%. Also in the RT-LAMP method without RNA extraction, the sensitivity of the technique was 85. 7%. Conclusion:  The LAMP-RT PCR technique is emerging as an appropriate alternative to the Real-Time RT-PCR method. This technique has basic advantages, such as constant temperature amplification, thermal cycle elimination, faster results, and potentially greater detection capacity. Therefore, the RT-LAMP-PCR can be used as a quick and cost-effective technique that can be employed in all areas