Patent DE-102009009228-A1 (DE)
Title: Agglutination-based detection of disease comprises adding substrate of buffer, measuring initial intensity of buffer, diluting blood sample with buffer, measuring reference intensity and originating test person with disease to diagnose
Inventor: WOLF GUENTHER
IPC: G01N33/483
Filed: 2009-02-17
Abstract: Agglutination-based detection of specific diseases through a blood test, comprises: adding a substrate quantity of a buffer solution; measuring the initial intensity (IA) of the buffer solution, diluting a blood sample with buffer solution; measuring the reference intensity (IO); storing the measurement data as vector (V1i), originating the test person with typical characteristics of the disease to be diagnosed; displaying the definite classified states of positive (pathological) and negative (healthy); and storing the result with the uncertain condition in a column vector (V2i). Agglutination-based detection of specific diseases through a blood test, which consists of one sample preparation, a measurement process and a data analysis with a classification, comprises: adding a substrate quantity of a transparent buffer solution, mixing, heating and subsequently irradiating by light; measuring the initial intensity (IA) of the transparent buffer solution in a first measuring step, diluting a blood sample with buffer solution and adding its partial quantity of the initial volume of the transparent buffer solution and mixing with it; measuring the reference intensity (IO) in a second measuring step and calculating the start extinction (EO) from the ratio of the intensities (IO) and (IA), where to balance different hematocrit values, the samples of the defined normal range of the optical density are verified, and correcting at range below- or above progressing to the normal range, measuring again the initial intensity (IA1) and subsequently removing again the excess of the blood sample buffer mixture, and by adding a specific lectin or antibody of the agglutination process, and thus initiating the measurement of the agglutination kinetics to the time (t1), recording a specific measurement value (I1), calculating the initial value of the extinction E1 of the agglutination of the ratio (I1) and (IA1), measuring all other measurement values with the intensities I(t) in time steps, from I(t) and (IA1), determining the current extinction E(t), terminating the measurement after a defined number of time steps; storing the measurement data as vector (V1i), where the vector (V1i) of the measurement value E(t) is evaluated by classifying and comparing with threshold values, and before the first initial operation, the evaluation to measure the neural networks for classification with at least one record is in the set P1 (p1, p2, p3) of patterns, originating the test person with typical characteristics of the disease to be diagnosed, and training a record of the set G (g1, g2, g3) with characteristics of healthy volunteers and initializing, assigning the vector (V1i) of the measurement progression E(t) after passing through a first neural network, to one of at least three classes, of which the positive state, a negative state and another uncertain state of the patient relative to the disease to be diagnosed represents; displaying the definite classified states of positive (pathological) and negative (healthy); and storing the result with the uncertain condition in a column vector (V2i), together with other column vectors, which are divided into parameter groups and contain estimated data from the history of the patient, and classifying, evaluating the class results, where the result of the evaluation is transferred, and carrying out the class release of at least three classes (positive, negative and uncertain) and displaying a reference for further investigations for the uncertain state. An independent claim is included for a device, for carrying out the above method, comprising a measuring device from a tempering- and sample receiving station, a spectrophotometer and an embedded-control-unit with specific storage units, where the tempering- and sample receiving station are assembled with a heater, a Peltier cooler and a rotating field generator with a quartz stabilized control system, which is provided for defined localization, and fixation of a transparent measuring cuvette and a recording, and a magnetic stirrer is inserted into the measuring cuvette, where the tempering- and sample receiving station are disposed via a metering device with three channels each with a micro-pump and a micro-valve for the injection or suction of buffer solution, blood sample buffer mixture and lectin, where a light source of the spectrophotometer is arranged, which irradiates the measuring cuvette during the measurement process, and the selective measurement value of a specific wavelength of the blood sample is sampled in time steps on the spectrophotometer, and is recorded in the storage unit of the spectrometer controller, which is the part of the embedded-control-unit, where in the embedded-control-unit and in the storage unit, the structure of a two-stage classification- and evaluation-network for classification, evaluation and storage of the classes of the agglutination process are stored, and in the structure of two-stage classification- and evaluation network, an initial classification network, consisting of a first neural network and a first evaluation circuit, and a second classification network, consisting of a second neural network, a second evaluation circuit and the XOR-elements for outputting the signals (Apa) for positive, (Ana) for negative and the output directly from the second evaluation circuit for the signal (Aua) for uncertain are arranged, where the second classification network is bound with the output of the class three of the first evaluation circuit and the column vectors, via a data- and control bus, the embedded-control-unit and the first classification network alternative to the transfer of the learning data stored in the embedded-control-unit of a learning data matrix or the data vectors (V1i) and are coupled to the transfer of control signals, and are stored in the storage of the embedded-control-unit data to issue instructions for further diagnostic studies of the release uncertain state.