Fecal occult blood latex-enhanced immunoturbidimetric kit

A latex-enhanced, immune-turbidimetric technology, applied in the field of clinical in vitro reagent testing, can solve the problems of missed detection, low sensitivity of colloidal gold method, and inability to quantitatively analyze the analyte, and achieves simple and convenient operation, easy automation, and detection stability. and good repeatability

Inactive Publication Date: 2018-07-24
天津市协和医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common immunological method is the colloidal gold method, which uses hemoglobin as the detection index. The sensitivity of the colloidal gold method is low, and the inability to perform quantitative analysis on the analyte is still a shortcoming of this method. The higher sensitivity is that the hemoglobin content exceeds 100ng / ml to show positive, and the test result is negative when the hemoglobin content is less than 100ng / ml, which causes missed detection to a certain extent

Method used

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  • Fecal occult blood latex-enhanced immunoturbidimetric kit
  • Fecal occult blood latex-enhanced immunoturbidimetric kit
  • Fecal occult blood latex-enhanced immunoturbidimetric kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of R1 reagent:

[0029] Glycine buffer 0.1M; PEG60002wt%; ProClin-300 0.1wt%; pH=7.4; the balance is water; weigh 7.507g glycine; PEG6000 20g; ProClin-300 1g; add 500ml of water to dissolve and mix; use 3M NaOH aqueous solution Adjust the pH to 7.4±0.1; add water to make the volume to 1L.

Embodiment 2

[0031] R2 reagent is made by the following method:

[0032] (1) Take 100mg polystyrene carboxyl microspheres with a particle size of 60nm (JSR company, carboxyl content 0.187meq / g), wash twice with 50mM MES buffer with pH=5.5, and use 50mM MES buffer with pH=5.5 Dilute the solution to 10ml;

[0033] (2) Add 3 mg of N-hydroxysulfosuccinimide to the 10 ml suspension obtained in step (1), mix well, and add 2 mg of 1-(3-dimethylaminopropyl)-3-ethyl carbon Diimine hydrochloride, react for 30min under dark stirring, 15000rpm, centrifuge for 45min, discard the supernatant;

[0034] (3) Resuspend the centrifugal product obtained in step (2) with 9ml of pH=8.0 boric acid buffer solution, sonicate it with ultrasound for 2 seconds at intervals of 2 seconds until the suspension is uniform, add 0.5mg of rabbit anti-human hemoglobin polyclonal The antibody was reacted for 3h under stirring, 15000rpm, centrifuged for 30min, and the supernatant was discarded.

[0035] (4) Add 10ml of blocking solut...

Embodiment 3

[0039] R2 reagent is made by the following method:

[0040] (1) Take 100mg polystyrene carboxyl microspheres with a particle size of 145nm (JSR company, carboxyl content 0.104meq / g), wash once with 50mM MES buffer with pH=5.5, and use 50mM MES buffer with pH=5.5 Dilute the solution to 10ml;

[0041] (2) Add 1 mg of N-hydroxysulfosuccinimide to the 10 ml suspension obtained in step (1), mix well, and add 2 mg of 1-(3-dimethylaminopropyl)-3-ethyl carbon Diimine hydrochloride, react for 15min under dark stirring, 15000rpm, centrifuge for 20min, discard the supernatant;

[0042] (3) Resuspend the centrifugal product obtained in step (2) with 9ml of pH=8.0 boric acid buffer solution, sonicate for 2 seconds at intervals of 2 seconds until the suspension is uniform, add 3.0mg rabbit anti-human hemoglobin polyclonal The antibody was reacted under stirring for 4h, 15000rpm, centrifuged for 15min, and the supernatant was discarded.

[0043] (4) Add 10ml of blocking solution to the centrifugal...

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Abstract

The invention discloses a fecal occult blood latex enhanced immunoturbidimetric kit comprising a R1 reagent, a R2 reagent and a human hemoglobin calibrator; the R1 reagent comprises a glycine buffer,PEG6000 and ProClin-300; the R2 reagent comprises a glycine buffer, rabbit anti-human hemoglobin-coated latex microspheres, bovine serum albumin, Tween-20, trehalose, ProClin-300; the latex microspheres in the reagent R2 have a particle size of 60 to 220 nm, and a hemoglobin antibody is rabbit anti-human hemoglobin polyclonal antibody, and is linked to the polystyrene latex microspheres by covalent coupling. Latex enhanced immunoturbidimetric detection results objectively reflect the amount of bleeding, the fecal occult blood latex enhanced immunoturbidimetric kit facilitates observing of changes in conditions of diseases; due to a full automatic operation mode, the detection process is more convenient and quick, and the fecal occult blood latex enhanced immunoturbidimetric kit can test inbatches, and is more suitable for screening experiments of large-scale populations.

Description

Technical field [0001] The invention relates to the field of clinical in vitro reagent testing, in particular to a fecal occult blood latex enhanced immune turbidimetric method detection kit. Background technique [0002] Colorectal cancer is one of the most common diseases today. Approximately 1.2 million patients worldwide are diagnosed with colorectal cancer each year, and more than 600,000 patients die of colorectal cancer directly or indirectly. In my country, the incidence and mortality of colorectal cancer are respectively the 3rd and 4th among malignant tumors, and the annual incidence rate is twice the world average, and the 5-year survival rate of colorectal cancer patients has been lingering for a long time. The low level is mainly due to the low proportion of early-stage cancers among the colorectal cancers admitted to most hospitals. Therefore, improving the level of early diagnosis is an important part of improving the survival rate of patients with colorectal cance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N33/531
CPCG01N33/721G01N33/531
Inventor 石牡丹王立凯张续
Owner 天津市协和医药科技集团有限公司
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