A traditional nested reverse transcription-polymerase chain reaction (RT-PCR) assay specific for eastern equine encephalomyelitis (EEE) virus was designed to multiplex with a previously described West Nile (WN) virus nested RT-PCR assay. Differentiation of EEE and WN was based on base pair size of the amplified product. One hundred fifty-seven mammalian and avian brain tissues were tested by EE...

This study evaluated and compared the performance of two real-time PCR assays using nested reverse transcription (RT)-PCR as the reference method. Among 117 nested RT-PCR-positive cases, the abTES DEN 5 qPCR kit detected 97.4% of dengue virus (DENV) infections, while the innuDETECT Dengue TwoStep assay did so for 44.4%. Sensitivity varied by infecting serotype and the stage of infection. The ab...

The spreading of viral RNA within Swiss Albino mice orally inoculated with coxsackievirus B4 E2 strain (CVB4 E2) was studied by using RT-PCR and semi-nested-RT-PCR methods. Viral RNA was detected in various organs: pancreas, heart, small intestine, spleen, thymus, and blood at various postinfectious (p.i.) times ranging from 8 hr to 150 days. Our results show that (i) outbred mice can be infect...

Background: Spermatogenesis associated 19 (Spata19) was introduced as a testis-specific gene that probably involved in spermatogenesis cell apoptosis. Therefore, this study aimed to investigate the effect of Spata19 inactivation on sperm count. Materials and Methods: We generated global knockout mice by CRISPR/Cas9 nickase technology. Disability validated three levels DNA, RNA, protein using PC...

Human rhinoviruses (HRVs) are the major cause of the common cold. HRVs were recently reclassified into the Enterovirus genus (HEV) in the Picornaviridae family. HRVs and other members of the HEV genus share many common features, including sense RNA genomes and partial nucleotide sequence identity. The aim of this study was to evaluate different HRV detection strategies. Samples from adults with...

Several methods for the detection of Borna disease virus (BDV) RNA have been reported, one being the reverse transcription-nested polymerase chain reaction (RT-nested PCR) method. However, due to the possibility of contamination of the cloned DNA in a reaction tube, false-positive results might be obtained by RT-nested PCR. To detect only BDV RNA without anxiety of contamination, we developed a...

bovine viral diarrhoea virus (bvdv) is an important pathogen of dairy cattle. in this study, bulk milk samples representing a total of 4105 milking cows, from 18 dairy cattle herds in the suburb of mashhad- iran, were tested for presence of bvdv by the use of a nested reverse transcription polymerase chain reaction (nested rt- pcr) assay. non of the cows in the herds had been vaccinated against...

A nested reverse transcriptase (RT) PCR assay detected mRNA of the salmonid pathogen Renibacterium salmoninarum in samples of RNA extracts of between 1 and 10 cells. Total RNA was extracted from cultured bacteria, Atlantic salmon (Salmo salar L.) kidney tissue and ovarian fluid seeded with the pathogen, and kidney tissue from both experimentally challenged and commercially raised fish. Followin...

Laboratory testing for dengue virus is used to confirm the diagnosis of dengue virus infection and to differentiate dengue from other febrile tropical illnesses. There are few data on the clinical use of reverse transcription-polymerase chain reaction (RT-PCR) for diagnosis of dengue virus infection. We prospectively evaluated 121 consecutive patients with possible dengue who had samples submit...