摘要
目的观察重组人肌酸激酶MM同工酶(CK-MM)在不同基质中的稳定性和互通性,筛选出适合的基质制备校准品用于CK测定的标准化。方法将重组酶分别置于自制类人血清基质、抗冻基质、仿有证参考物配方基质及人血清基质中,用常规测定方法观察重组酶的稳定性;分别采用参考方法、常规方法(罗氏试剂和中生试剂)测定不同基质中的重组酶活性和不同浓度的人血浆总CK活性,观察重组酶在不同方法间的互通性。结果重组酶在自制类人血清基质和仿有证参考物配方基质中-20℃冻存可稳定25d和23d,在抗冻基质中至少可稳定240d。自制类人血清基质和仿有证参考物配方基质中重组酶在参考方法与常规方法间有互通性,人血清基质和抗冻基质中的低值重组酶样品在两方法间具有互通性。结论自制基质和有证参考物配方基质中的重组酶具有较好的稳定性和互通性,初步具备了校准品的基本特点。本研究为进一步研制CK测定的参考品,包括校准品和定值控制品提供了思路。
Objective To investigate the stability and comparability of recombinant human muscle creatine kinase isozyme（CK-MM） in different matrix, and to find out a suitable matrix as a reference material for application of CK measurement system. Methods The stability of the recombinant CK-MM was observed by measuring its catalytic activity in self-made matrix ,anti-freeze additive matrix,the certified reference material （CRM） matrix and human serum matrix respectively. For understanding the commutability among different measurement system total CK catalytic activity in various concentration of human serum and the catalytic activity of recombinant CK-MM in different matrix specimen were measured with IFCC recommended reference method and routine reagent produced by Roche and Zhongsheng Diagnostic Company respectively. Results The recombinant CK-MM was stable for 25 days in the self-made matrix,23 days in CRM matrix and at least 240 days in antifreeze additive matrix stored at - 20 ℃. The recombinant enzyme protein in self-made matrix and CRM matrix was commutable. Low concentrations of recombinant CK-MM in human serum and in anti-freeze additive matrix were also commutable. Conclusion The recombinant CK-MM had good stability and commutability in either self-made matrix or CRM matrix. It might be used as a candidate reference material for the determination of total creatine kinase in serum.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2007年第5期333-335,共3页
Chinese Journal of Clinical Laboratory Science
