Optimization and Preliminary Methodology Study of in vitro Fresh Human Whole Blood Detection Method for Pyrogen-related Factor IL-1β
10.6039/j.issn.1001-0408.2016.19.17
- VernacularTitle:体外新鲜人血法检测热原相关因子IL-1β的条件优化及初步方法学研究
- Author:
Wenjia WANG
;
Zhiming CHEN
;
Zhihui PANG
;
Haishun FANG
;
Huahong HE
;
Wei LI
- Publication Type:Journal Article
- Keywords:
Human fresh blood;
Pyrogen;
Interleukin 1β;
in vitro test;
Qingkailing injection;
Ginaton injection
- From:
China Pharmacy
2016;27(19):2644-2646,2647
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish the detection method for pyrogen-related factor interleukin 1β(IL-1β)through optimiz-ing detection condition,and to conduct preliminary methodology study. METHODS:The in vitro fresh human whole blood detec-tion method was used. The bacterial endotoxin standard solution(5,2,0.8,0.32 EU/ml)were added into diluted blood;using di-luted RPMI 1640 as blank control,the content of IL-1β in blood sample was determined by ELISA after incubation. The relation-ship of the addition of different attenuants(RPMI 1640 culture,sterilized normal saline)and fetal bovine serum,final dilution vol-ume fraction(40%,20%,10%and 8.3%)and storage duration(2,5,6,8,26 h)with the contents of endotoxin IL-1βwere in-vestigated,and related coefficient and detection limits were calculated. Different dilution times of Qingkailing injection and Ginaton injection samples and interference solutions were added into diluted blood to detect their recovery. RESULTS:The results indicated that RPMI 1640 media and 40% diluted blood was more sensitive(detection limit was 0.128 EU/ml,r=0.993);while the addition of fetal bovine serum didn’t influence the results. The detection limits of blood sample storied at 4 ℃ for 26 h were 0.128 EU/ml (r>0.990). When Qingkailing injection and Ginaton injection were diluted 10,32 and more times,the detection method was not interfered and the recovery ranged 68%-118%. CONCLUSIONS:Established in vitro fresh human whole blood detection method can be used for the detection of pyrogen,and provides trial evidence for the pyrogen detection of TCM injection.
