ABSTRACT
Procalcitonin (PCT) is the precursor of calcitonin related to the severity of human bacterial infection. We made a test strip by coupling anti-PCT to quantum dot, in order to develop a highly sensitive and convenient PCT testing product. The anti-PCT titer had reached 10⁷ because of the stability by coupling anti-PCT with quantum dot. The detecting linear range of the experiment was 0.15 to 120 μg/L, the sensitivity was 0.007 μg/L, the recovery range was 91% to 113%, and the intra- and inter-assay coefficient of variation was less than 8%. Comparing the homemade fluorescence-detected test strip with PCT ELISA kit on sale, we got accurate results which could mostly accomplish the test of clinical samples.
ABSTRACT
We expressed 17-hydroxysteroid dehydrogenase10 (17β-hsd10) recombinant protein, prepared anti-17β- hsd10 polyclonal antibodies and established sandwich enzyme linked immunosorbent assay (ELISA) test for detection of 17β-hsd10. RT-PCR was used to get the gene of 17β-hsd10 of mouse liver, and a prokaryotic protein expression system pET 15b-17β-hsd10/Escherichia coli BL21 (DE3) which induced with isopropyl-1-thio-β-galactopyranoside (IPTG) for recombinant protein expression was constructed subsequently. The target protein purified using His-Binding-resin column was used to immunize BALB/c mice and rabbits, serum total IgGs from immunized animals were purified by ammonium sulfate precipitation method. We established a Double-antibody Sandwich enzyme linked immunosorbent assay about 17β-hsd10 using the two antibodies we prepared. We got the concentration of 1.5 mg/mL of 17β-hsd10 protein with molecular weight of 29.5 kDa, and polyclonal antibodies from mouse and rabbit with the tite 1.25 x 10(4) and 2.5 x 10(4) respectively. The concentration of 0.1 g/mL of 17β-hsd10 can be detected by the Double-antibody Sandwich ELISA we established, and the assay was sensitive and specific. It can be widely used in clinical and experimental study.