Nitto Boseki Co., Ltd.

National Institute of Agrobiological Sciences

Shinshu University

Ryo Kojima

Yoshiro Sato

Katsuhiro Katayama

Takeshi Nagasawa

Toshihide Miura

Kenta Noda

Iwao Kiyokawa

Yoshikiyo Sasagawa

Natsuki Sato

Hideki Sezutsu

Keiro Uchino

Isao Kobayashi

Toshiki Tamura

Yuji Arimatsu

Fumio Nomura

Kazuyuki Sogawa

Wataru Kikuchi

Kiyotaka Fujita

Chie Satokawa

Akiko Takekawa

Hirotoshi Tanaka

Yasunori Okazaki

Kaduho Ashizawa

Momoe Sato

Satoshi Inoue

Satoshi Arai

Disclosed is a test instrument for measuring an analyte in a liquid sample by a noble metal colloid aggregation measurement method. The test instrument involves a reaction chamber in which at least the liquid sample is to be reacted with a reagent, wherein the reagent is adhered on at least a part of a surface constituting the reaction chamber in a dried state, and the reagent enables the measurement of the analyte by a noble metal colloid aggregation measurement method. The test instrument additionally involves a supply section for supplying the liquid sample and a flow path for delivering the liquid sample that has been supplied to the supply section to the reaction chamber, wherein the liquid sample that has been supplied to the supply section is delivered to the reaction chamber through the flow path to cause the liquid sample to be brought into contact with the reagent that has been adhered in a dried state, thereby producing a difference in pressure between the supply section and the reaction chamber for the purpose of dispersing the reagent in the liquid sample. When the test instrument is used, the measurement based on an absorbance at a visible region can be achieved, the analyte can be measured accurately within a short time, and the measurement suitable for a POCT field can be achieved.

Test instrument for measuring analyte in sample by an aggregation assay using a metal colloid and using a reagent attached in a dry state in a reaction chamber, and method for measuring analyte using same

Pentameric CRP is produced at a high efficiency by transferring DNA, which encodes monomeric CRP, into a silkworm to thereby construct a transgenic silkworm and then collecting and purifying pentameric CRP that is produced by the transgenic silkworm constructed above.

DNA encoding canine monomeric CRP and expression vector containing the DNA

Disclosed is an immunoassay method whereby a 5.9 kDa peptide which results from the degradation of the &#x3b1;-E chain and &#x3b1; chain of human fibrinogens and which is used as a peptide marker for diagnosing hepatic disease can be specifically assayed in a biological sample containing contaminating peptides by bringing antibodies that recognize the N terminal of said peptide marker and antibodies that recognize the C terminal of said peptide marker into contact with said peptide marker, forming immune complexes of said peptide marker and the two antibodies, and immunoassaying the obtained immune complexes.

5.9 kDa peptide immunoassay method

In the case of assaying the level of complex AB in a sample which likely contains the complex AB composed of substance A with another substance B, the complex is assayed by the competitive homogeneous assay method with the use of reagents including a reagent containing partner C specifically binding to substance A, a reagent containing partner D specifically binding to substance B, a reagent containing fine particles carrying substance A or an analog thereof and substance B or an analog thereof, and a reagent containing partner C specifically binding to substance A and partner D specifically binding to substance B. Thus, the complex in the sample can be easily assayed. The above method is applicable to general-purpose biochemical automatic analyzers.

Method of assaying complex and kit to be used therefor

Pentameric CRP-producing transgenic silkworm

In measuring phosphoric acid by the use of an enzyme cycling system using a dehydrogenase together with a thio-NADP, a thio-NAD, a reduced thio-NADP or a reduced thio-NAD as a coenzyme, phosphoric acid is measurable in a wide concentration range from a low concentration to a high concentration by measuring phosphoric acid after previous removal of free phosphoric acid in reagent components for the measurement.

Method for measuring phosphoric acid

By using an antibody which is covalently bonded to a water soluble polymer as an antibody to be used in a competitive immunoagglutination assay in a homogeneous system, a protein antigen at a high concentration can be accurately assayed in an undiluted system without resorting to dilution.

Method of assaying antigen and kit to be used therein

A method for determination of calcium in a sample derived from a living body, by the reaction of calcium in the sample with Chlorophosphonazo-III or a compound analogous thereto in the presence of vanadate ions, followed by determination of calcium in the sample on the basis of an optical change caused by the reaction product, is excellent in various respects as follows: it is free from the problem of absorption of carbonic acid gas caused by a high pH; it is free from the problem of use of a toxic reagent containing arsenic; it makes it possible to subject a large number of samples to determination in a short time because it can be applied to an autoanalyzer; and it permits determination in a wide range because of low sample blank values.

Reagent for determination of calcium and determination method

A two reagent-type kit for assaying GPT by acting GPT on L-alanine and &agr;-ketoglutarate in the presence of pyridoxal phosphate, converting the resulting pyruvate into lactate with L-lactate dehydrogenase (LDH) in the presence of reduced nicotinamide adenine dinucleotide (NADH) and measuring GPT based on a decrement of NADH, the GPT assay kit comprising a first reagent and a second reagent, one of which contains pyridoxal phosphate, L-alanine, &agr;-ketoglutarate, LDH and NADH and the other reagent contains no pyridoxal phosphate but contains L-alanine. The GPT assay kit is stable over a long period of time.

Reagent for GPT assay

There is provided an enzymatic method of rapid quantitative assay for 1,5-anhydroglucitol which is also applicable to an automated analysis device. The enzymatic method of quantitative assay for 1,5-anhydroglucitol is characterized by using a pyranose oxidase derived from,No. 52 as a pyranose oxidase. According to the assaying method of the present invention, assay for 1,5-anhydroglucitol in a specimen can be performed using an automated analysis device, results in rapid assay and improved accuracy.

Method of quantitative assay for 1,5-anhydroglucitol

By making a bilirubin oxidase derived from the genus Pleurotus to act on a living body fluid sample in the presence of at least one of a cationic surfactant such as alkyltrimethylammonium salt, etc. and a nonionic surfactant such as polyoxyethylenealkyl ether, etc., only bilirubin non-.delta. fractions of the living body fluid sample can be oxidized, while keeping a bilirubin .delta. fraction remaining as unreacted. Thus, the bilirubin non-.delta. fractions and .delta. fraction of the living body fluid sample can be exactly quantitatively determined by action of the bilirubin oxidase on the sample.

Method for assaying bilirubin .delta. fractions

A method of measuring urea nitrogen on the basis of the ultraviolet absorption of urease-GLDH by a reaction rate method, which method not only permits accurate measurement of a sample containing a high concentration of urea nitrogen but also permits accurate measurement of a sample containing polyols such as mannitol without suffering the influence of the polyols, the method comprising hydrolyzing urea in a sample with urease, reacting glutamate dehydrogenase (GLDH) with ammonia formed by the hydrolysis, in the presence of .alpha.-ketoglutaric acid (.alpha.-KG) and reduced-type nicotineamide adenine dinucleotide (NADH) or reduced-type nicotineamide adenine dinucleotide phosphate (NADPH), and measuring the reduced-type nicotineamide adenine dinucleotide (NADH) or reduced-type nicotineamide adenine dinucleotide phosphate (NADPH) for a decrease rate to measure urea nitrogen derived from the urea, and characterized in that a sulfhydryl compound is co-present with the urease.

Kit for measuring urea nitrogen

Method for measuring bilirubin

There is provided a method for determination of direct bilirubin which comprises the steps of contacting a bilirubin oxidase with a sample suspected of containing bilirubin; and, measuring direct bilirubin in the sample by optical changes of the sample, characterized in that bilirubin oxidase is allowed to act in the presence of a indirect bilirubin reaction inhibitor selected from a thiocyanate ion, a hydrazide, reduced nicotinamide adenine dinucleotide, reduced nicotinamide adenine dinucleotide phosphate and a potassium ion of 100 mM to 800 mM. By completely avoiding interference of indirect bilirubin, direct bilirubin is selectively and precisely determined using a reagent kit in the form of solution. The method and reagent of the invention are safe and free of environmental pollution due to unnecessary waste liquid treatment.

Method for determination of direct bilirubin and reagent therefor

A method of measuring the total ketone body in a sample comprising converting acetoacetic acid in the sample to 3-hydroxybutyric acid in advance and subsequently converting the whole 3-hydroxybutyric acid including 3-hydroxybutyric acid existing in the sample originally to acetoacetic acid to lead the reduction of nicotinamide adenine dinucleotide according to the action of 3-hydroxybutyrate dehydrogenase and a sample reagent thereof is disclosed. According to this invention an assay of the total ketone body in samples can be carried out conveniently, highly precisely and quickly.

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Koriyama, Japan

Ryo Kojima