[Development of one step RT-PCR technique for detection of H7 subtype avian influenza]

Sheng Wu Gong Cheng Xue Bao. 2009 Nov;25(11):1658-63

Authors: Bao H, Wang X, Tao Q, Cai D, Wang F, Chen H

According to 45 hemagglutinin (HA) gene sequences of H7 subtype of avian influenza virus (AIV), a pair of specific oligonucleotide primers was designed. We developed one step RT-PCR for detecting AIV subtype H7. Sensitivity to detection of allantoic fluid by one step RT-PCR reached 10(5.5) EID50/mL and detection of swab samples reached 10(3) EID50/mL. We simultaneity detected the tissue and swab samples infected with H7 subtypes AIV by one step RT-PCR and virus isolation method. The results showed that the sensitivity of the assay gave an excellent correlation with the conventional virus isolation method. H1-H15 subtypes of avian influenza and other avian diseases were detected by the one step RT-PCR. The results showed the assays were high specific, without cross-reaction with other subtypes or other avian diseases. Development of one step RT-PCR will provide effective technical support for the rapid diagnosis and surveillance of molecular epidemiology of AIV subtype H7.

PMID: 20222464 [PubMed - in process]

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Potent inhibitors of furin and furin-like proprotein convertases containing decarboxylated P1 arginine mimetics.

J Med Chem. 2010 Feb 11;53(3):1067-75

Authors: Becker GL, Sielaff F, Than ME, Lindberg I, Routhier S, Day R, Lu Y, Garten W, Steinmetzer T

Furin belongs to the family of proprotein convertases (PCs) and is involved in numerous normal physiological and pathogenic processes, such as viral propagation, bacterial toxin activation, cancer, and metastasis. Furin and related furin-like PCs cleave their substrates at characteristic multibasic consensus sequences, preferentially after an arginine residue. By incorporating decarboxylated arginine mimetics in the P1 position of substrate analogue peptidic inhibitors, we could identify highly potent furin inhibitors. The most potent compound, phenylacetyl-Arg-Val-Arg-4-amidinobenzylamide (15), inhibits furin with a K(i) value of 0.81 nM and has also comparable affinity to other PCs like PC1/3, PACE4, and PC5/6, whereas PC2 and PC7 or trypsin-like serine proteases were poorly affected. In fowl plague virus (influenza A, H7N1)-infected MDCK cells, inhibitor 15 inhibited proteolytic hemagglutinin cleavage and was able to reduce virus propagation in a long-term infection test. Molecular modeling revealed several key interactions of the 4-amidinobenzylamide residue in the S1 pocket of furin contributing to the excellent affinity of these inhibitors.

PMID: 20038105 [PubMed - indexed for MEDLINE]