新生儿苯丙酮尿症（PKU）荧光筛查系统的研究
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摘要
苯丙酮尿症(PKU)是一种由于人体正常染色体缺陷,造成的新生儿体内苯丙氨酸代谢紊乱性疾病。如果在婴儿出生早期没有得到及时诊断和治疗,在发育过程中,高浓度的苯丙氨酸会严重损害婴儿的神经系统,将导致病儿不可逆的智力障碍,所以对其早期筛查尤为重要。目前国内还没有独立知识产权的PKU检测诊断仪器,本课题的主要目的就是研究一套适合我国国情的PKU筛查系统。
本文主要论述以下几个内容:
1.推演了苯丙氨酸与茚三酮的反应机理,在此基础上设计了干血片荧光法苯丙氨酸检测试剂盒的制作方案,提出了增强和稳定荧光的方法。并通过大量临床对比试验,完成了国产新生儿PKU筛查试剂盒的研制。
2.选取新型超亮紫外发光二极管作激发光源,采用不同直径的石英光纤和玻璃光纤束构成Y型传光光路,并利用自聚焦光纤对光线进行准直,使用光电倍增管进行检测,设计成新型光纤荧光新生儿PKU筛查光学系统,并对设计原理进行了理论推演和实验验证。该系统具有结构简单、体积小、重量轻、灵敏度高等特点。
3.采用平行双轨道结构设计,利用两个步进电机驱动,四个传感器定位,设计了标准96孔微孔板精密扫描系统,并兼有对样品混合震荡和恒温反应功能。
4.采用嵌入式PC结构进行PKU检测装置的硬件电路设计,完成对光电检测信号的预处理、A/D转换、扫描驱动、初始定位、微孔板震荡和恒温控制等功能。并基于WINDOWS操作系统,利用VC语言编写控制和数据处理分析软件。系统智能化程度高、便于维护和升级、统计分析功能强并可直接网上上报数据。
临床试验表明:本系统性能稳定,操作简单,结果准确,成本低,满足临床筛查要求,为我国PKU的大批量筛查提供了新的快速检测手段。本课题是天津市社会发展基金资助项目,目前已经通过天津市科委项目验收。
Phenylketonuria(PKU) is a genetic disease as a result of chromosome abnormality, which can cause the metabolization disorder of phenylalanine. If Left untreated, the disorder would damage the neural system greatly and cause nonreversible mental retardation as a result of the accumulation of phenylalanine . Therefore, it is of great importantance to diagnose and treate PKU as early as posible for nowborns. Unfortunately, there is no domestic instrument specially designed for PKU screening at the present time. The aim of this project is to develop a fluorescence detection system for neonatal phenylketonuria screening.
Main points discussed in this article are listed as follows:
1.Deduced the reaction mechanism of phenylalanine and ninhydrin, based on which we designed a designed a new reagent which used in phenylalanine measurement. Found a new method to aggrandize and stabilize fluorescence and also we devoloped the first demestic reagent box for PKU screening through many clinic experiments.
2.Using a high luminance ultraviolet LED for excitation source, a“Y”type fiber which consists of quartz and Glass fiber with different diameter for conduction of light, an autofocus fiber for focalization, and a photo multiplier tube for measurement. Designed a new optical system for neonatal phenylketonuria screening and demonstrate the feasibility of it in theory. Experiments show that this optical system is sensitive, simple, small and light.
3.Adopted a double-parallel-rail frame, and used two marching electromotor for driving, four sensor for locating and standard 96-tiny hole board system for scanning, which at the same time has the ability to mix specimens and keep constant temperature.
4.Adopted embedded PC frame for the designing of the hard ware circuits of the PKU detecting system which has multiple functions such as the preprocessing of electrical and optical signal, A/D converting, scanning driving, tiny-hole board shaking and constant temperature keeping, etc. Designed a data processing software using VC++ based on WINDOWS. The system has the advantages in the following aspects: highly intellectualized, easy maintenance and upgrading, strong statistical and analyzing capability, direct access to the internet, etc.
Clinic experiments indicate that the system is stable, easily operated, accurate, and with low cost, which meets the demand of clinic application. This system provides a new method for mass PKU detecting. The project is supported by Tianjin Social Development fund and has been accepted by Tianjin Scientific Committee.
本文主要论述以下几个内容:
1.推演了苯丙氨酸与茚三酮的反应机理,在此基础上设计了干血片荧光法苯丙氨酸检测试剂盒的制作方案,提出了增强和稳定荧光的方法。并通过大量临床对比试验,完成了国产新生儿PKU筛查试剂盒的研制。
2.选取新型超亮紫外发光二极管作激发光源,采用不同直径的石英光纤和玻璃光纤束构成Y型传光光路,并利用自聚焦光纤对光线进行准直,使用光电倍增管进行检测,设计成新型光纤荧光新生儿PKU筛查光学系统,并对设计原理进行了理论推演和实验验证。该系统具有结构简单、体积小、重量轻、灵敏度高等特点。
3.采用平行双轨道结构设计,利用两个步进电机驱动,四个传感器定位,设计了标准96孔微孔板精密扫描系统,并兼有对样品混合震荡和恒温反应功能。
4.采用嵌入式PC结构进行PKU检测装置的硬件电路设计,完成对光电检测信号的预处理、A/D转换、扫描驱动、初始定位、微孔板震荡和恒温控制等功能。并基于WINDOWS操作系统,利用VC语言编写控制和数据处理分析软件。系统智能化程度高、便于维护和升级、统计分析功能强并可直接网上上报数据。
临床试验表明:本系统性能稳定,操作简单,结果准确,成本低,满足临床筛查要求,为我国PKU的大批量筛查提供了新的快速检测手段。本课题是天津市社会发展基金资助项目,目前已经通过天津市科委项目验收。
Phenylketonuria(PKU) is a genetic disease as a result of chromosome abnormality, which can cause the metabolization disorder of phenylalanine. If Left untreated, the disorder would damage the neural system greatly and cause nonreversible mental retardation as a result of the accumulation of phenylalanine . Therefore, it is of great importantance to diagnose and treate PKU as early as posible for nowborns. Unfortunately, there is no domestic instrument specially designed for PKU screening at the present time. The aim of this project is to develop a fluorescence detection system for neonatal phenylketonuria screening.
Main points discussed in this article are listed as follows:
1.Deduced the reaction mechanism of phenylalanine and ninhydrin, based on which we designed a designed a new reagent which used in phenylalanine measurement. Found a new method to aggrandize and stabilize fluorescence and also we devoloped the first demestic reagent box for PKU screening through many clinic experiments.
2.Using a high luminance ultraviolet LED for excitation source, a“Y”type fiber which consists of quartz and Glass fiber with different diameter for conduction of light, an autofocus fiber for focalization, and a photo multiplier tube for measurement. Designed a new optical system for neonatal phenylketonuria screening and demonstrate the feasibility of it in theory. Experiments show that this optical system is sensitive, simple, small and light.
3.Adopted a double-parallel-rail frame, and used two marching electromotor for driving, four sensor for locating and standard 96-tiny hole board system for scanning, which at the same time has the ability to mix specimens and keep constant temperature.
4.Adopted embedded PC frame for the designing of the hard ware circuits of the PKU detecting system which has multiple functions such as the preprocessing of electrical and optical signal, A/D converting, scanning driving, tiny-hole board shaking and constant temperature keeping, etc. Designed a data processing software using VC++ based on WINDOWS. The system has the advantages in the following aspects: highly intellectualized, easy maintenance and upgrading, strong statistical and analyzing capability, direct access to the internet, etc.
Clinic experiments indicate that the system is stable, easily operated, accurate, and with low cost, which meets the demand of clinic application. This system provides a new method for mass PKU detecting. The project is supported by Tianjin Social Development fund and has been accepted by Tianjin Scientific Committee.
引文
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