Avian Influenza (AI) is caused by type A influenza virus belonging to the family orthomyxoviridae, which is classified into 16 HA and 9 NA subtypes based on two surface glycoprotein’s haemagglutinin (HA) and neuraminidase (NA). In the present study we did identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR) during the first outbreaks of AI in India duri...

One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RT-PCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis...

RT-PCR assays for detection of BRSV, based on four different sets of primers were optimized and evaluated for their sensitivity and specificity. Primers used in this study were specific for genes encoding three BRSV proteins, nucleoprotein N and glycoproteins F and G. Our results indicated that RT-PCR with primers B7:B8 for G protein was the most efficient in detecting BRSV. Starters B7:B8 reac...

We have developed a combinatorial polymerase chain reaction (PCR) procedure to identify four major species of the genus Brucella simultaneously. Four pairs of primers targeting the genes encoding a cell surface protein (BCSP31) and outer membrane proteins (omp2b, omp2a and omp31) were prepared. PCR using these primers gave rise to specific patterns of amplification for each Brucella spp. examin...

In order to improve simultaneous detection and identification of Helicobacter genus in general and Helicobacter pylori specifically and reduce the number of amplifications needed, we established a multiplex PCR. In this study, two pairs of primers: Hcom1 and Hcom2 specific for Helicobacter genus, Hicd1 and Hicd2 specific for Helicobacter pylori species were used. To determine the sensitivity of...

Quantitative PCR can often be improved by conducting the amplification with nested primers. First, fewer nonspecific amplification products, which could otherwise interfere with quantitation, are produced. Often, nonspecific products can be eliminated. In these cases, relatively simple nonspecific detection techniques are suitable for quantitation. In addition, nested primer PCR provides intrin...

wild grasses are the most important primary feedstuffs which are susceptible to contamination with toxigenic fungi belonging to aspergillus spp. in order to explore diversity of aspergillus species associated with the inflorescences of gramineous weeds, infected inflorescences were collected from wild grasses in western parts of iran. fifty-six aspergillus isolates were obtained from all diseas...