CAJ | 학술논문

目的：构建一种以磁珠为标记物的，用于血型检测的可视化蛋白质芯片。方法用人免疫球蛋白G （IgG）及磁珠标记的抗人IgG进行琼脂糖浓度、点样浓度、温度和固定时间等实验条件的摸索。根据优化的条件，经过铺片、点样、固定、封闭等程序构建ABO血型检测蛋白质微阵列，加入血液样品进行反应，最后用磁珠标记的抗体进行检测。结果通过条件的优化发现，在1．0％的琼脂糖基片表面，以100μg/m L的蛋白质稀释液点样，之后在24℃固定8 h ，能够得到更优的信号强度。检测不同稀释度的抗原抗体，正定型效价可达到1∶8192，反定型可达到1∶4096。检测14例临床样品，结果样点清晰、肉眼可见，且结果判定与试管法一致性达100％。结论成功构建了基于磁珠的血型检测可视化蛋白质微阵列。
목적：구건일충이자주위표기물적，용우혈형검측적가시화단백질심편。방법용인면역구단백G （IgG）급자주표기적항인IgG진행경지당농도、점양농도、온도화고정시간등실험조건적모색。근거우화적조건，경과포편、점양、고정、봉폐등정서구건ABO혈형검측단백질미진렬，가입혈액양품진행반응，최후용자주표기적항체진행검측。결과통과조건적우화발현，재1．0％적경지당기편표면，이100μg/m L적단백질희석액점양，지후재24℃고정8 h ，능구득도경우적신호강도。검측불동희석도적항원항체，정정형효개가체도1∶8192，반정형가체도1∶4096。검측14례림상양품，결과양점청석、육안가견，차결과판정여시관법일치성체100％。결론성공구건료기우자주적혈형검측가시화단백질미진렬。
Objective To construct a visual blood typing protein microarray based on magnetic bead .Methods The optimal experiment conditions ,such as agarose concentration ,temperature ,immobilization concentration and time ,were explored on glass-slide with human IgG and anti-human IgG antibody coated magnetic bead .Based on the optimal conditions ,an ABO blood typing protein microarray was established by coating ,printing ,immobilizing and blocking .Then the blood samples were added to the microarrays ,and detected with antibody-conjugated magnetic beads .Results When the class slide was coated with 1% agarose ,and spotted with the protein probes (100 mg/mL in printing buffer) on the fabricated substrates and incubated in a humid chamber at 24 ℃ for 8 h ,better results could be obtained .The forward test titer was up to 1∶819 2 ,and the reverse test was 1∶409 6 by detecting different dilu-tion rate of samples with the arrays .14 clinical specimens were tested with the arrays and tube methods ,and the re-sults were same .Each positive spot on the arrays was strong and visible easily to the naked eye .Conclusion A sys-tem of magnetic bead-based protein array for ABO blood group was successfully constructed .