ABSTRACT
The paper described the rationality of traditional and modern application of prepared decoction pieces of herbal medicine on basis of application, statistics and comparison analysis of three forms of drugs of traditional Chinese herbal pieces prepared for decoction, prepared decoction pieces in small packing and granules; and illustrated different opinions correlative to the three forms of drugs; put forward the counter-measures and proposals for the problems facing the traditional Chinese herbal pieces for decoction; the paper stated clearly that the traditional Chinese herbal pieces for decoction should not be replaced, instead, the viewpoint and the reasons on its application must be holding on; and the trend of development and expectations of the Chinese herbal pieces for decoction were predicted as well.
Subject(s)
Dosage Forms , Drugs, Chinese Herbal , Medicine, Chinese Traditional , Methods , Phytotherapy , MethodsABSTRACT
<p><b>OBJECTIVE</b>To prepare and characterize monoclonal antibodies (mAb) and polyclonal antibodies against nucleocapsid (N) protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) and to establish antibodies-based sandwich ELISA for detecting N protein of SARS-CoV, which might apply to early diagnosis of patients with SARS-CoV infection.</p><p><b>METHODS</b>BALB/c mice were immunized with purified recombinant N protein of SARS-CoV for producing mAbs, and New Zealand white rabbits were immunized for producing polyclonal antibodies. The identification of antibodies was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. Capturing and detecting antibodies were selected by pairing the mAbs and polyclonal antibodies one by one and an antibodies-based sandwich antigen capture ELISA was used for detecting N antigen of SARS-CoV.</p><p><b>RESULTS</b>Nine mAbs and hyperimmune rabbit polyclonal antibodies, specifically against SARS-CoV nucleocapsid protein were obtained. Using paired ELISA assay, three mAbs N1E8, N8E1 and N10E4 were selected as capturing antibody and rabbit polyclonal antibodies as detecting antibody then triple antibodies-based sandwich ELISA was established following horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G. The recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml with this assay. When tested with 420 serum specimens from serologically confirmed SARS patients, the positive rates of serum N protein were 90.1%, 23% and 0%, in which sera collected from 1 to 10 days, 11 to 20 days and beyond 21 days respectively after the onset of symptoms. The specificity of the assay was 99.86% in 715 control serum specimens. There was no cross-reaction with other respiratory viruses and coronaviruses.</p><p><b>CONCLUSION</b>Specific and high affinity mAbs and rabbit polyclonal antibodies were obtained. By paired and optimized sandwich ELISA, a sensitive and specific antigen capture ELISA was established for detecting N antigen of SARS-CoV, which might apply to early diagnosis, source tracing and epidemiological studies of SARS.</p>