WO2022247523A1 - 一种用于结直肠癌早期诊断的生物标志物组合物及应用 - Google Patents
一种用于结直肠癌早期诊断的生物标志物组合物及应用 Download PDFInfo
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Abstract
Description
Claims (13)
- 一种结直肠癌生物标志物组合物,其特征在于,所述生物标志物组合物包括口炎消化链球菌Peptostreptococcus stomatis,具核梭状杆菌Fusobacterium nucleatum,微小微单胞菌Parvimonas micra,双歧杆菌Bifidobacterium。
- 一种检测结直肠癌的试剂盒,其特征在于,所述试剂盒包括权利要求1所述生物标志物组合物的量化试剂,所述量化试剂用于量化权利要求1所述的生物标志物组合物中的每一组分。
- 如权利要求2所述的试剂盒,其特征在于,所述量化试剂采用的原理为实时定量PCR法。
- 如权利要求3所述的试剂盒,其特征在于,所述量化试剂包括用于扩增权利要求1所述生物标志物组合物的引物组,所述引物组包括用于扩增权利要求1所述生物标志物组合物每一组分的引物对。
- 如权利要求4所述的试剂盒,其特征在于,所述引物组的核苷酸序列如SEQ ID NO.1-8所示。
- 如权利要求5所述的试剂盒,其特征在于,所述量化试剂还包括用于扩增总菌的引物对。
- 如权利要求6所述的试剂盒,其特征在于,所述用于扩增总菌的引物对的核苷酸序列如SEQ ID NO.9-10所示。
- 如权利要求7所述的试剂盒,其特征在于,所述量化试剂还包括实时定量PCR反应试剂。
- 如权利要求2-8任一所述的试剂盒,其特征在于,所述试剂盒还包括用于检测粪便中人血红蛋白的隐血检测试剂。
- 如权利要求9所述的试剂盒在结直肠癌早期诊断中的应用,其特征在于,所述试剂盒用于检测结直肠癌时的步骤为:检测权利要求1所述的生物标志物组合物中每一组分在样本中的丰度以及粪便隐血值,再通过回归方程判断检测结果。
- 如权利要求10所述的试剂盒在结直肠癌早期诊断中的应用,其特征在于,所述试剂盒用于检测结直肠癌时,包括以下步骤:(1)粪便隐血值测定:采用粪便隐血检测试剂盒测定受试者粪便样品的隐血结果值X 0,其中隐血结果为阳性时X 0=1,隐血结果为阳性时X 0=0;(2)以受试者粪便基因组DNA为模板,采用试剂盒中的量化试剂分别检测每个生物标志物的扩增值Ct 标志物,和总菌的扩增值Ct 总菌;(3)根据公式(Ⅰ)分别确定样本中生物标志物组合物中每一组分在总菌中的相对含量Ct值,Ct=lg(2^(Ct 标志物-Ct 总菌))(Ⅰ);(4)根据回归方程计出算Y值,并将Y值带入公式(Ⅱ)计算P值,其中,e为自然常数,(5)结果判定:当P>0.5,结直肠癌诊断结果为阳性;当P≤0.5,结直肠癌诊断结果为阴性。
- 如权利要求11所述的应用,其特征在于,所述步骤(4)中的回归方程为式(Ⅲ)所示,Y=A+β 0X 0+β 1X 1+β 2X 2+β 3X 3+β 4X 4(Ⅲ);其中,X 0为隐血结果值,X 1为口炎消化链球菌Peptostreptococcus stomatis的Ct值,X 2为具核梭状杆菌Fusobacterium nucleatum的Ct值,X 3为微小微单胞菌Parvimonas micra的Ct值,X 4为双歧杆菌Bifidobacterium的Ct值;A和β 0-β 4均为常数,A和β 0-β 4由临床实验数据获得。
- 如权利要求12所述的应用,其特征在于,所述步骤(4)中的回归方程包括式(Ⅳ)-式(Ⅷ)中的任一种:Y=-3.9583+3.1015X 0-0.6478X 1-0.3980X 2+0.2912X 3-0.5130X 4(Ⅳ);Y=-2.965+2.7005X 0-1.2309X 1-0.6054X 2+0.3328X 3-0.2359X 4(Ⅴ);Y=-3.099+1.9907X 0-2.0011X 1-0.3290X 2+0.5476X 3-0.6620X 4(Ⅵ);Y=-3.771+4.0089X 0-0.9983X 1-0.4482X 2+0.1288X 3-0.4917X 4(Ⅶ);Y=-2.9043+2.8920X 0-0.5471X 1-0.1049X 2+0.2281X 3-0.6194X 4(Ⅷ)。
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