F-specific (F+) RNA phages are recommended as indicators of fecal contamination and the presence of enteric viruses and as viral surrogates to elucidate the resistance of viruses to adverse conditions or to assess the effectiveness of inactivating processes. Reverse transcription (RT)-PCR methods have been used to detect, quantify, or identify subgroups of F+ RNA phages. However, these methods ...

The E. coli flagellar types H1 and H12 show a high serological cross-reactivity and molecular serotyping appears an advantageous method to establish a clear discrimination between these flagellar types. Analysis of fliC H1 and fliC H12 gene sequences showed that they were 97.5% identical at the nucleotide level. Because of this high degree of homology we developed a two-step real-time PCR detec...

step. The presence of Vent DNA polymerase and PCR products other than the intended DNA linker, however, did not seem to interfere with subsequent reactions because ligations using both gel-purified and unpurified PCR products yielded a similar number of transformants. Eight transformants were tested, all of which contained the PCR product as an insert. The yield was considered to have high effi...

To rapidly distinguish Canine distemper virus (CDV), canine parvovirus (CPV), and canine kobuvirus (CaKoV) in practice, a one-step multiplex PCR/RT-PCR assay was developed, with detection limits of 102.1 TCID50 for CDV, 101.9 TCID50 for CPV and 103 copies for CaKoV. This method did not amplify nonspecific DNA or RNA from other canine viruses. Therefore, the assay provides a sensitive tool for t...

Purpose: Fusobacterium nucleatum is an opportunistic pathogen involved in periodontal diseases, extraoral infections, and colorectal cancer. necrophorum causes a variety of necrotic infections. F. are classified into five two subspecies, respectively. Conventional identification methods were technically hard to distinguish each subspecies species accurately. The purpose the present study was de...

during 1387-86, total number of 120 blood samples gathered from sheep with bluetongue-like clinical sign. the samples collected from seropositive regions .after separating serum, they were evaluated by competitive elisa for detecting ab against vp7 antigen. the full length of s7 gene (1156 bp) amplified by one step rt-pcr .in this method two sets specific primers, targeting 3? and 5?ends of s7 ...

Two-dimensional (2-D) gene scanning (TDGS) is a method for mutation detection based on the electrophoretic separation of PCR-amplified DNA fragments according to size and base pair sequence. The use of denaturing gradient gel electrophoresis (DGGE) as the second separation step provides virtually 100% sensitivity, while the 2-D format allows the inspection of multiple gene fragments simultaneou...

Fungal infections are frequently observed in living-donor liver transplantation (LT) and are correlated with the clinical outcome. Early diagnosis of fungus infection is difficult because of the low positive rate of blood culture and the poor specificity of the (1-3)-D-glucan assay for fungi. A polymerase chain reaction (PCR)-based method was developed for the early diagnosis of fungal infectio...

A simple and effective modified ethanol precipitation-based protocol is described for the preparation of DNA from ancient human bones. This method is fast and requires neither hazardous chemicals nor special devices. After the powdering and incubating of the bone samples Dextran Blue was added as a carrier for removing the PCR inhibitors with selective ethanol precipitation. This method could e...