A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus

Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

Expression of three S100A calcium-binding proteins in human gastric cancer and non-cancerous mucosa as detected by QRT-PCR / 解放军医学杂志

Objective To verify the authors' previous cDNA micro-array results and to further investigate the moleculer mechanisms of gastric cancer. Methods Quantitative real-time RT-PCR was employed to detect the expressions of three S100A calcium-binding genes in 22 fresh surgical samples of gastric tumor tissue and non-cancerous mucosa from the same patients. Results The transcription level of S100A2 in primary cancer lesion was elevated in 80% of samples when compared with matching non-neoplastic mucosa (P=0.018) and the average up-regulation level was 10.78 fold. 55% of cancer lesions showed higher transcription level of S100A4 than their adjacent non-neoplastic mucosa, the average up-regulation level was 2.31 fold. S100A6 transcription level was higher in 74% (P=0.01) of primary cancer lesion with an 2.25 fold up-regulation than the adjacent non-neoplastic mucosa. After rectified by ?_2-microglobulin, the relative expression levels of S100A2, S100A4 and S100A6 were 2.83?10~ -4 , 6.44?10~ -2 and 0.41, respectively. According to the Spearman correlation coefficient analysis there were significant positive correlations between S100A2 and S100A4, and S100A2 and S1006 (P value were 0.00 and 0.017, respectively). Conclusion The changes in S100A2 and S100A6 genes may be an early event in a majority of gastric cancer patients, while S100A4 may be associated with the infiltration of gastric cancer. Further study on the three genes might be helpful for understanding the nature of gastric carcinoma.