| Objective:On the basis of a blood coagulation sensor detection system construction of using piezoelectric quartz crystal (PQC), after detecting conditions of the biosensor being optimized, a series of experimental researches using the sensors have been used to detect the plasma fibrinogen and coagulation factor, in order to explore the feasibility of clinical application.Methods:1. Technology of fine processing was used firstly to make the quartz crystal, and the electrodes were silvered on the surface. Then the mucilage-style micro-sensors-array was prepared on the organic glass with small holes. And we developed the self-oscillatiion circuits and the PESA software (version 2.0) to record and analyze the frequency. In this way, a blood coagulation array sensor detection system of using piezoelectric quartz crystal was constructed by connecting with a computer.2. The construction of the micro-sensors-array system of detecting plasma fibrinogen and coagulation factor: Firstly, the detection conditions, such as temperature, humidity, and time, were measured. Secondly, the standard curves were drawn. Thirdly, the sensitivity and linearity scope of the detection were observed. Then the detection results of clinical samples were compared with those of the STAGO fully automatic analysis instrument, in order to explore the feasibility of detecting the clinical samples.3. The clinical research of piezoelectric quartz crystal blood coagulation biosensor diagnosis array: Firstly, Prothrombin time (PT), Activate partial thromboplastin time (APTT), Thrombin time (TT), Fibrinogen (FIB) and coagulation factor (FⅧ) of the clinical samples were detected. Then the results of 40 clinical samples detected by the sensors were compared with the detection results by the STAGO fully automatic analysis instrument. Theresearch provides the foundation for the clinical application and popularization of the piezoelectric quartz crystal blood coagulation analysis instrument. Results:1.1 quartz crystal sensors: The sensor with a basic frequency of 10MHz was constructed by AT-cut with the quartz crystal diameter of 8.5 mm, and it had silver film with the diameter of 4.0mm on the surface. This quartz crystal can start to vibrate easily, with a stable vibration frequency and a high sensitivity. It is suitable for the detections in blood coagulation reaction.1.2 The self-excitation oscillation circuit invented by ourselves has strong vibratory ability at liquid phase, and its stability of frequency also can meet the request of the experiment.1.3 The PESA software(version 2.0) developed by ourselves is applying to record and analyze frequency had a lot of functions, such as setting the initial parameters automatically, recording the lively frequency variation, drawing the tendency scheme of the frequency variation automatically, and analyzing the results. The data of collection and analysis could be transmitted to the computer automatically.1.4 The coating of PPE solution on the surface of the crystal reduced the frequency variation of the crystal oscillator in the liquid and detection error. It also improved the stability of the detection frequency (±lMHz at gaseous phase, ±2MHz at liquid phase) and the sensitivity of the sensors.1.5 The sample-feed slot designed by our labtory reduced the influence of frequency while the sample being fed.1.6 The method of mucilage-style let it easy to make a testing pool, and it can detect a singular item or multiple items in same time.2. At the condition of the temperature 34±2°C, the humidity 48±2%, the Fibrinogen and coagulation factor Vfll could be detected quickly with our piezoelectric quartz crystal blood coagulation sensors. The sensitivity and linearity scope of the detection results were fit with the results detected by the standand clinic methods very well. It shows that our piezoelectric quartz crystal blood coagulation sensors can be used in clinic successfully.3. While the sensors were used in the detection of clinical samples at the conditions given(the temperature 34±2°C, the humidity 48±2%), the clinical samples detection resultsof Prothrombin time (PT), Activate partial thromboplastin time (APTT), Thrombin time (TT), Fibrinogen (FIB) and coagulation factor (FVI)coincided with the detecting results by the STAGO fully automatic analysis instrument, and the relational coefficients were 0.935 > 0.931> 0.936, 0.947, 0.946 respectively. Conclusion:1. When piezoelectric quartz crystal sensor array oscillates at liquid phase, special optimization is processed by the self-oscillating circuit and hence stable oscillation is realized.2. PESA (version 2.0) is a kind of new-style software.lt can record and analyze the resonance frequency of piezoelectric quartz crystal sensor array in live.3. The coating of PPE solution on the surface of the crystal reduces the frequency variation of the crystal oscillator in the liquid and detection error. It also improves the stability of the detection frequency (±lMHz in gaseous phase, ±2MHz in liquid phase) and the sensitivity of the sensors. The problems such as the influence of silver film quickly being oxidated to the detection results and the limited number of the prepare sensors are solved.4. It is easy to make the mucilage-style sensors, and the sensors are very cheap. It can detect a singular item or multiple items in one time.5. The design of the sample-feed slot reduces the influence of frequency while the sample being fed.6. The method using the piezoelectric quartz crystal blood coagulation biosensor is a quicker, the more sensitive and more accurate method after the colorimetry and the magnetic bead method. It is also a cheap and simple method. Besides, it is a new method that can detect every step in the reaction processes on line and in live. Moreover, the start and the end point of the reaction can be decided easily.7. The temperature and humidity have great influence on the coagulation reaction, and it is always a pivotal problem during the detecting process.8. The results of Fibrinogen and coagulation factor FVffl detected by the piezoelectric quartz crystal blood coagulation biosensor coincide with those detected by the STAGO fully automatic analysis instrument.9. After a series of detections to the clinical samples were performed, we can concludethat the results of the coagulation blood indexes detected by the piezoelectric quartz crystal blood coagulation biosensor are accurate. It is a simple, cheap, quick, sensitive method, and it is very valuable for clinical application.
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