Author Archives: Dan Frost

Expression and identification of eCTLA4-N/GP fusion proteins

The expressing of eCTLA4-N fusion protein was firstly determined in prokaryotic system. Supernatant and cell lysates of E.coli DH5α transformed with pET30a/eCTLA-S or pET30a/S plasmid were analyzed by SDS-PAGE (Figure 1A). Neither eCTLA4-N fusion protein nor HTNV N was expressed into supernatant (Figure 1A, lane 1 and lane 3). Instead, eCTLA4-N fusion protein was shown […]

December 22, 2012 – 5:44 pm | By Dan Frost | Posted in Other | Comments (0)

Serologic assays

HTNV N-specific IgG antibodies in mice sera were determined, by enzyme-linked immunosorbent assay (ELISA) in [26,27]. 96-well microtiter plates (Costar, USA) coated with 100 μl of purified recombinant N protein of HTNV strain A9 at a concentration of 1 μg/ml as described previously. Hantaan virus glycoproteins specific IgG antibodies were evaluated by IFA using insect […]

December 21, 2012 – 3:28 pm | By Dan Frost | Posted in Diseases | Comments (0)

Cells and viruses

The 293T, Vero E6 cells and Baby hamster kidney cell (BHK) cells were purchased from ATCC (ATCC number: CRL-1586) and cultured in Dulbecco modified Eagle medium (DMEM) supplemented with 10% heat-inactivated fetal calf serum, 100 U of penicillin, and 100 μg of streptomycin per ml at 37 with 5% CO2. HTNV strain 84Fli, isolated from […]

December 20, 2012 – 6:12 pm | By Dan Frost | Posted in Diseases | Comments (0)

Fusion with extracellular domain of cytotoxic T-lymphocyte-associated-antigen 4 leads to enhancement of immunogenicity of Hantaan virus DNA vaccines in C57BL/6 mice

Background Hantaan virus (HTNV) is the causative agent of the most severe form of a rodent-borne disease known as hemorrhagic fever with renal syndrome (HFRS). A safe and effective HTNV vaccine is needed. Vaccination with DNA constructs expressing fused antigen with bioactive factors, has shown promising improvement of immunogenicity for viral agents in animal models, […]

December 19, 2012 – 6:56 pm | By Dan Frost | Posted in Diseases | Comments (0)

The development of a rapid SYBR Green I-based quantitative PCR for detection of Duck circovirus

This report describes a one-step real-time polymerase chain reaction assay based on SYBR Green I for detection of a broad range of duck circovirus (DuCV). Align with all DuCV complete genome sequences and other Genus Circovirus download from the GenBank (such as goose circovirus, pigeon circovirus), the primers targets to the replicate gene of DuCV […]

November 10, 2011 – 4:16 pm | By Dan Frost | Posted in Infections | Comments (0)

Risk assessment for HCV infection. Part 2

The incidence and prevalence of HCV among dialysis patients varies markedly from country to country and among dialysis centers within a single country. HCV prevalence is much higher in developing countries as compared to developed world. First paper in Pakistan in this line of literature was published in 1999 and till yet six different reports […]

November 4, 2011 – 11:58 am | By Dan Frost | Posted in Hepatitis | Comments (0)

RNA Isolation, PCR amplification and Detection

The striking genetic heterogeneity of the RNA genome of HCV is well recognized. On the basis of molecular relatedness, HCV is classified into 11 major genotypes: 1 through 11, among which first six are major player of infection globally. On the basis of phylogenetic analysis, over 80 subtypes and minor variants referred to as “quasispecies […]

November 4, 2011 – 6:11 am | By Dan Frost | Posted in Hepatitis | Comments (0)

Rising burden of Hepatitis C Virus in hemodialysis patients

High prevalence of Hepatitis C virus (HCV) has been reported among the dialysis patients throughout the world. No serious efforts were taken to investigate HCV in patients undergoing hemodialysis (HD) treatment who are at great increased risk to HCV. HCV genotypes are important in the study of epidemiology, pathogenesis and reaction to antiviral therapy. This […]

November 2, 2011 – 7:10 pm | By Dan Frost | Posted in Hepatitis | Comments (0)

Risk assessment for HCV infection

This study involved 384 patients treated with HD and was screened for anti HCV and tested for HCV RNA and HCV genotypes during January 2010-April 2010. The mean age of the patients was 40.9 ± 5.54 years (age rang15-75 years). 244 were male (mean age 40.4 ± 8.4) and 140 were female (age range 41.7 […]

October 27, 2011 – 7:38 pm | By Dan Frost | Posted in Hepatitis | Comments (0)

Generation of double-stranded DNA using long oligos and Taq polymerase

20 pmol of each paired oligo was incubated with 2 U of Taq DNA polymerase (Promega, Madison, USA), 1X reaction buffer, 2.5 mM MgCl2, and 0.4 mM of dNTPs. The reaction mix was incubated at 94°C for 30 seconds followed by 55°C for 30 seconds and 72°C for 30 seconds for 5 cycles. After the […]

September 25, 2011 – 3:51 am | By Dan Frost | Posted in Other | Comments (0)