ABSTRACT
Objective To develop a double-antibody sandwich ELISA for determining the concentration of nucleoprotein (NP) of rabies virus in various products of rabies vaccine.Methods The purified rabies antibodies from a rabbit were used to coat microwell plates. Horseradish peroxidase-conjugated anti-NP monoclonal antibody was used to probe the NP bound to coated antibodies.The assay was used to quantitatively determine the concentration of NP of rabies virus.Results The results showed that the coefficient of linear correlation was higher than 0.97.The optimal linear range was 0.000625~0.01 IU/ml and the detection limit was 0.000625 IU/ml. The recovery rate was 102~109% and the coefficient of variation was only 7.2%~9.4%.No cross reactions were observed with bovine serum,bovine serum albumin,ovalbumin,refined solution of influenza vaccine,encephalitis B vaccine,and hepatitis A vaccine.Conclusions The results indicated that the assay is specific,sensitive,accurate,reproducible,and stable,and could be suitable for quantitative determination of different rabies vaccine's processes products.
ABSTRACT
Objective To study the early diagnosis of cutaneous T-cell lymphoma(CTCL).Methods DNA from39formalin-fixed,paraffin-embedded tissues of24cases of CTCL,2cases of dubious lymphoma,and4cases with non-specific erythroderma,was amplified by polymerase chain reaction(PCR),with special primers of TCR V?1-8,V?9,and J?1/?2to analyze T-Cell receptor?gene clonal rearrangement(TCR-GR clonal).The specimens from30patients with non-specific dermatitis were taken as negative control.Results Clonal of TCR V?1-8was demonstrated in38of39paraffin-embedded specimens.V?9-GR was shown in37of39specimens TCR?-GR was shown especially in13patients with MF of early stage,2patients with dubious CTCL,and4patients with non-specific erythroderma,as well as in6of30patients with histiologically nonspecific dermatitis which suggested that these6cases were"clonal dermatitis".Conclusion Clonal TCR?-GR may be detected in patients with early CTCL,even when the histologic findings are not unequivocally diagnostic.
ABSTRACT
Objective To establish an efficient, simple, low cytotoxicity and cheap transfaction system. Methods We have used the cationic polymer polyethylenimine(PEI) to study transient transfection in MCF-7 cells by testing different conditions, including cell concentrations, DNA concentrations, the ratio of PEI nitrogen to DNA phosphate(PEI-N∶DNA-P) and the time of cells grown in serum-free culture together with PEI-DNA complex. Results The optimized cell concentrations were 2?10 5 cells seeded per well in 24-well dishes 18-24?h before transfection. The DNA concentrations and ratio of PEI-N∶DNA-P are very important for optimal transfection and they affect each other. For 1??g DNA per well, the optimal PEI-N∶DNA-P is about 33∶1, however, as for 4??g DNA, it is 9∶1. The best time of cells grown in serum-free culture together with PEI-DNA complex is about 5-7?h.Conclusion With optimized conditions, we can establish an efficient, simple, low cytotoxicity and cheap transfection system by using PEI.