血色原微量分光法血痕确证试验

摘要

目的：探讨血色原微量分光法作为血痕确证试验的可行性。方法用离心分离的检材浸洗液代替检材，与改良高山试剂进行血色原反应，用微量分光光度计测定吸收光谱，判断检材是否含血。在优化参数的基础上，用倍比稀释的粗制人血红蛋白测定方法的灵敏度，用常见疑似血液/血斑，以及不同保存时间、不同基质的血斑测定方法的特异性和检材适应能力。结果用血色原微量分光法，仅血液（斑）具有415nm、525nm和555nm 三个吸收峰组成的特异光谱，与常见疑似血液/斑没有交叉。反应2min，上样2.5μL，稀释1000倍的人血可稳定获得三峰光谱。通过延长浸洗时间、设置基质对照，10年陈旧血纱、有色布上的血斑等均可有效确证。三个吸收峰的高度与检液的血含量呈正相关，且525nm和555nm的吸光度变化趋势一致（y=0.5232x+0.0274，R2=0.9971）。结论血色原微量分光法灵敏度高、特异性强，快速简便，可纳入DNA检验流程之中，在实际检案中有较好的应用前景。替代传统结晶法用于教学，更符合当前学生技能和意识的培养要求。%Objective To explore the feasibility of hemochrome identification by micro spectrophotometer. Methods We use dip lotion by Centrifugation, reaction with modiifed takayama reagent,measured absorption spectra with a micro spectrophotometer to determine whether samples contain blood. On the basis of optimized parameters, we use diluted human hemoglobin to measure sensitivity of the method; using suspected blood / blood spots, and different storage time and matrix of blood spots to measure speciifcity and samples of adaptability. Result With micro spectroscopic method of hemochromogen,only blood (spot) has speciifc spectral consisting of three absorption peaks at 415,525 and 555nm and no cross suspected common blood / spot.Reaction 2 min, sample volume2.5ul, 1000-fold diluted human blood stably obtains primordial spectrum. By prolonging the immersion time, set the vehicle control, 10 years old blood gauze. Blood stains in colored cloth can be effective detective.. The height of absorption peaks and blood content were significantly corelated, and the change in absorbance at 525 and 555nm consistent trend (y=0.5232x+0.0274, R2=0.9971). Conclusion Micro spectroscopic method of hemochromogen has high sensitivity and speciifcity, quick and easy, can be incorporated into the DNA testing process. There is a good prospect in the actual seizure case. Instead of the traditional crystallization method for teaching, training requirements more in line with the skills and awareness for current students.