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Kit for detecting premature rupture of membrane by taking Axl as detection index and preparation method thereof

A technology for detecting kits and cloning antibodies, which is applied to measuring devices, instruments, scientific instruments, etc., to achieve the effect of improving accuracy

Active Publication Date: 2013-03-27
ORIGISSAY BIOLOGICS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The relationship between Axl and pregnancy has not been reported

Method used

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  • Kit for detecting premature rupture of membrane by taking Axl as detection index and preparation method thereof
  • Kit for detecting premature rupture of membrane by taking Axl as detection index and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] In this embodiment, the following process steps are used to prepare the premature rupture of membranes detection kit with Axl as the detection index:

[0046] (1) Preparation of multi-well plates coated with Axl monoclonal antibody

[0047] The Axl monoclonal antibody was diluted to a concentration of 10 μg / ml with carbonate buffer solution of pH=9, and then the Axl monoclonal antibody dilution was added to each well of the 96-well plate in an amount of 100 μl / well, Coating at 4°C for 12 hours; after the expiration of the coating time, add bovine serum albumin solution with a concentration of 1g / 100ml in the amount of 120μl / well to each well on the multi-well plate and carry out blocking reaction at 37°C, the reaction time is 1 hour; after the blocking reaction, wash the multi-well plate with pH=7.2 and 0.05mol / l phosphate buffer solution, and obtain the multi-well plate coated with Axl monoclonal antibody after the unreacted substances on the multi-well plate are remov...

Embodiment 2

[0053] In this example, the premature rupture of membranes detection kit prepared in Example 1 is used to detect the opposite sample.

[0054] 1. Sample preparation

[0055] (1) Standard sample

[0056] The Axl protein standard item equipped in the kit described in Example 1 is set as the standard sample containing the highest concentration of Axl, named as standard sample 1, and the concentration of its Axl is 6000pg / ml, and then used in the kit described in Example 1 The prepared sample diluent dilutes the Axl protein standard into standard samples with Axl concentrations of 2000pg / ml, 666.7pg / ml, 222.2pg / ml, 74.07pg / ml, 24.69pg / ml, and 8.23pg / ml respectively, They are named Standard Sample 2, Standard Sample 3, Standard Sample 4, Standard Sample 5, Standard Sample 6, and Standard Sample 7 in sequence, and there are 7 standard samples in total.

[0057] (2) Sample

[0058] Taking outpatients and inpatients of the West China Second Hospital of Sichuan University as the exp...

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Abstract

The invention discloses a kit for detecting premature rupture of membrane by taking Axl as a detection index, which comprises a perforated plate coated with Axl monoclonal antibody, biotin-marked Axl monoclonal antibody (Biotin-AxlAb) detection liquid, avidin-horse radish peroxidase combined with biotin-marked Axl monoclonal antibody, a chromogenic substrate 3',3',5,5'-tetramethylbenzidine, and an Axl protein standard substance. A preparation method for the kit comprises the following technological steps of: (1) preparing the perforated plate coated with the Axl monoclonal antibody; (2) preparing the biotin-marked Axl monoclonal antibody (Biotin-AxlAb) detection liquid; and (3) preparing avidin-horse radish peroxidase, the chromogenic substrate 3',3',5,5'-tetramethylbenzidine, and the Axl protein standard substance.

Description

technical field [0001] The invention relates to the application of Axl receptor tyrosine kinase monoclonal antibody, a sandwich enzyme-linked immunosorbent assay kit and a preparation method for detecting premature rupture of fetal membranes (Premature Rupture of Fetal Membrane, PROM for short) in pregnant women. Background technique [0002] Rupture of Fetal Membrane (ROM) may occur at any time during pregnancy, and rupture of fetal membranes before labor is called premature rupture of membranes (PROM). The incidence of PROM after term (37 weeks of gestation) is about 10%; the incidence of PROM before term (before 37 weeks of pregnancy) is 2-3.5%. Premature rupture of membranes is the main factor causing prenatal and postpartum complications in obstetric patients, and the main reason for premature birth and neonatal admission to the intensive care unit. Because medical personnel have to balance the prolongation of gestational weeks with the risk of intrauterine and materna...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 胡怀忠关祥乾向洪发周容
Owner ORIGISSAY BIOLOGICS TECH
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