臍帯血や胎便などに含まれる微量なCRP定量法の確立と血中SAA定量法による初乳中のSAA測定に関する検討

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Quantitative method of C-reactive protein (CRP) progress greatly, and high-sensitivity assay is firmly established in the present well. We have already made clear that CRP is included in human colostrum by using EVIA method, one of the high-sensitivity assays. However, amniotic fluid and feces might include less amount of CRP which can not determine using this method, and therefore we aimed establishment of more sensitive determination by enzyme-1inked immunosorbent assay (ELISA). The minimum detectable concentration (sensitivity) of CRP was 0.05 ng/ml in contrast with EVIA having been 20 ng/ml. CRP could be measured in the range of 0.1 to 10 ng/ml. Three breast milk samples (mean CRP concentrations 6, 54, and 108 ng/ml) were run in duplicate in eight independent assays. The sample showed good inter-assay precision with %CV of 7.9, 4.8, and 3.3 respectively. The proposed ELISA demonstrated an excellent correlation with commercial CRP assay performed on an EVIA by analyzing 63 milk samples, with correlation coefficient above 0.96 and slop of approximately 1. Using the ELISA, we confirmed that CRP was contained in all of nine meconium and in eight-elevenths case of the amniotic fluid. The meconium CRP levels were ranged from 0.2 to 1, 030 ng/ml (mean 442±181), and the CRP levels in eight amniotic fluids were 0.2-720 ng/ml (mean 370±345). We examined a condition when we used conventional methods to measure whether serum amyloid A protein (SAA) was contained within colostrum. The concentration of SAA included in cryopreserved whole milk and whey obtained from the same puerpera was each 2, 294 ng/ml, 1, 548 ng/ml. Both a whole milk and whey lasted for a long term, and the alteration was not observed by the SAA concentration when we stored breast milk in a frozen state of -30℃. As a result, that human colostrum contained SAA and that the whey which treated to remove fat decreased to about 70% concentration of a whole milk were shown. This is the first report that showed that CRP is included in amniotic fluid and meconium, and showed that conventional method is effective to explore SAA in human breast milk and to measure its concentration.ELISAC-reactive proteinColostrumHuman whey serum amyloid A proteinAmniotic fluid
東北大学の論文
2004-07-31

臍帯血や胎便などに含まれる微量なCRP定量法の確立と血中SAA定量法による初乳中のSAA測定に関する検討

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