WO2021232713A1 - 一种新型冠状病毒IgG抗体的酶联免疫法检测试剂盒 - Google Patents
一种新型冠状病毒IgG抗体的酶联免疫法检测试剂盒 Download PDFInfo
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- the preparation of the anti-FITC antibody ELISA plate includes the following steps: dilute the anti-FITC antibody with 0.02-0.05M phosphate buffer to 1 ⁇ 5 ⁇ g/mL, add it to the transparent plastic ELISA plate at the same time, and coat it at 37°C 2-4 hours; discard the liquid in the well, wash the plate with pH 7.4 PBS buffer, then add phosphate buffer containing 0.5-2% BSA by mass to block the microtiter plate, and block at 37°C for 2-4 hours; discard Dry the liquid in the hole for 4-12 hours at 37°C; put it into an aluminum foil bag, add a desiccant, seal and label it, and store it at 2-8°C.
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Abstract
Description
样本编号 | 性别 | 年龄 | 诊断 | OD | S/CO | IgG判断 | 核酸 |
EG-0016 | 男 | 19 | 排除 | 0.071 | 0.71 | - | - |
EG-0031 | 女 | 40 | 排除 | 0.0399 | 0.40 | - | - |
EG-0034 | 女 | 48 | 排除 | 0.0244 | 0.24 | - | - |
EG-0077 | 男 | 12 | 排除 | 0.043 | 0.43 | - | - |
EG-0108 | 女 | 53 | 排除 | 0.0965 | 0.97 | - | - |
EG-0109 | 男 | 71 | 排除 | 0.0433 | 0.43 | - | - |
EG-0154 | 男 | 9 | 排除 | 0.0125 | 0.13 | - | - |
EG-0155 | 男 | 65 | 排除 | 0.072 | 0.72 | - | - |
EG-0185 | 女 | 53 | 排除 | 0.0906 | 0.91 | - | - |
EG-0186 | 男 | 18 | 排除 | 0.0154 | 0.15 | - | - |
EG-0211 | 女 | 31 | 排除 | 0.0027 | 0.03 | - | - |
EG-0212 | 女 | 50 | 排除 | 0.0065 | 0.07 | - | - |
EG-0239 | 女 | 29 | 排除 | 0.0067 | 0.07 | - | - |
EG-0245 | 男 | 54 | 排除 | 0.0428 | 0.43 | - | - |
EG-0288 | 女 | 1 | 排除 | 0.0764 | 0.76 | - | - |
EG-0289 | 男 | 55 | 排除 | 0.0603 | 0.60 | - | - |
样本编号 | 性别 | 年龄 | 诊断 | OD | S/CO | IgG判断 | 核酸 |
EG-0003 | 男 | 40 | 确诊 | 0.7984 | 7.98 | + | + |
EG-0004 | 男 | 46 | 确诊 | 0.9003 | 9.00 | + | + |
EG-0039 | 男 | 53 | 确诊 | 0.5679 | 5.68 | + | + |
EG-0042 | 男 | 51 | 确诊 | 0.3286 | 3.29 | + | + |
EG-0244 | 女 | 53 | 确诊 | 1.6752 | 16.75 | + | + |
EG-0246 | 女 | 60 | 确诊 | 1.5137 | 15.14 | + | + |
EG-0372 | 女 | 55 | 确诊 | 0.8239 | 8.24 | + | + |
EG-0375 | 男 | 1 | 确诊 | 1.0142 | 10.14 | + | + |
EG-0422 | 女 | 32 | 确诊 | 0.4869 | 4.87 | + | + |
EG-0425 | 男 | 57 | 确诊 | 0.5585 | 5.59 | + | + |
EG-0429 | 女 | 60 | 确诊 | 0.4948 | 4.95 | + | + |
EG-0431 | 男 | 52 | 确诊 | 1.2827 | 12.83 | + | + |
EG-0437 | 男 | 45 | 确诊 | 1.2299 | 12.30 | + | + |
EG-0439 | 男 | 35 | 确诊 | 1.5481 | 15.48 | + | + |
Claims (7)
- 一种新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:包括抗FITC抗体酶标板、FITC标记新型冠状病毒抗原、辣根过氧化物酶标记鼠抗人IgG单克隆抗体;其中,FITC标记新型冠状病毒抗原中的新型冠状病毒抗原包括合成多肽SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4;且4种合成多肽的质量比为1:1:1:1。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:所述FITC标记新型冠状病毒抗原的制备包括以下步骤,1)将FITC与合成多肽按摩尔比1:(1-3)进行混合,避光37℃反应6-12h;2)将上述步骤1)完成的标记液用0.01-0.05M PBS于2-8℃透析24h,加入等体积甘油,-20℃保存。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:还包括TMB显色剂A液和B液、以及终止液;所述终止液为浓硫酸、盐酸、氢氧化钠中的一种。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:还包括稀释液,Tris 1-8g/L;MES 2-5g/L;NaCl 5-20g/L;丙三醇20-50mL/L;海藻糖5-20g/L;Tween-20 1-5mL/L;ProClin TM300 0.5-2.5mL/L;硫酸庆大霉素2-5mL/L;pH值8.0±0.2。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:FITC标记新型冠状病毒抗原、辣根过氧化物酶标记鼠抗人IgG单克隆抗体的工作浓度均为0.01~0.5μg/mL。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:抗FITC抗体酶标板的制备包括如下步骤,将抗FITC抗体用0.02-0.05M磷酸盐缓冲液稀释至1~5μg/mL,同时加入到透明塑料酶标板中,37℃包被2-4小时;弃去孔内液体,用pH 7.4PBS缓冲液洗板,然后加入含质量浓度0.5-2%BSA的磷酸盐缓冲液封闭微孔板,37℃封闭2-4小时;弃去孔内液体,甩干后于37℃烘干4-12小时;装入铝箔袋,加入干燥剂,封口,贴标签,储存于2~8℃。
- 根据权利要求1所述的新型冠状病毒IgG抗体的酶联免疫法检测试剂盒,其特征在于:辣根过氧化物酶标记鼠抗人IgG单克隆抗体的制备方法如下,A、辣根过氧化物酶活化:1)配制5-10mg/mL HRP溶液;2)配制10-20mg/mL过碘酸钠NaIO 4溶液;3)将步骤1)和步骤2)配制溶液按体积比1:(1-3)混匀,2-8℃避光反应0.5-2h;4)配制浓度为10-40μL/mL的乙二醇水溶液,与步骤3)配制的溶液以相同体积混合,常温避光反应10-30min,活化即完成;B、辣根过氧化物酶标记鼠抗人IgG单克隆抗体:1)将待标记原料装入透析袋中,用0.02-0.1M pH8.5-10的碳酸盐缓冲液,透析0.5-2h;2)将标记原料与活化的HRP按质量比1:(1-5)进行混合,之后用0.02-0.1M碳酸盐缓冲液于2-8℃透析20-24h,期间换液2-3次;3)配制浓度为2-5mg/mL的NaBH 4水溶液,按1mgHRP加80μL配制好的NaBH 4水溶液的比例进行混合,并于2-8℃避光反应1-2h;4)将上述步骤3)完成的标记液用0.01-0.05M PBS于2-8℃透析20-24h,加入等体积甘油,-20℃保存。
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CN111551715B (zh) * | 2020-05-18 | 2022-04-22 | 天津博奥赛斯生物科技股份有限公司 | 一种新型冠状病毒IgM抗体的酶联免疫法检测试剂盒 |
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