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Premature rupture of membrane (PROM) detection kit using ICAM-1 as examination index and preparation method

A technology of ICAM-1 and detection kits, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as the preparation method of premature rupture of membranes detection kits that have not yet been discovered, and achieve the effect of improving accuracy

Active Publication Date: 2013-05-22
ORIGISSAY BIOLOGICS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although in the third trimester of pregnancy (gestational weeks 28-40), the ICAM-1 level in the amniotic fluid of pregnant women is increased by the expression of ICAM-1 on monocytes in the fetal membranes and amniotic fluid and the activation of neutrophils, but currently There is no report on the detection kit for premature rupture of membranes and its preparation method using ICAM-1 as the detection index

Method used

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  • Premature rupture of membrane (PROM) detection kit using ICAM-1 as examination index and preparation method
  • Premature rupture of membrane (PROM) detection kit using ICAM-1 as examination index and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0045] In this embodiment, the following process steps are used to prepare the premature rupture of membranes detection kit with ICAM-1 as the detection index:

[0046] (1) Preparation of a multi-well plate coated with ICAM-1 monoclonal antibody

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

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 to be tested

[0055] (1) Standard sample preparation

[0056] The ICAM-1 protein standard item equipped in the kit described in Example 1 is set as the standard sample containing the highest concentration of ICAM-1, named as standard sample 1, and the concentration of its ICAM-1 is 10ng / ml, then use the example 1 The sample diluent provided in the kit dilutes the ICAM-1 protein standard substance to ICAM-1 concentrations of 5 ng / ml, 2.5 ng / ml, 1.25 ng / ml, 0.63 ng / ml, 0.32 ng / ml, respectively. The standard samples of ml and 0.16ng / ml are named as 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 Un...

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Abstract

The invention discloses a premature rupture of membrane (PROM) detection kit using an intercellular adhesion molecular (ICAM)-1 as an examination index. The kit comprises a perforated plate coated with an ICAM-1 monoclonal antibody, biotin-labeled ICAM-1 monoclonal antibody (biotin-ICAM-1Ab) examination fluid, aidin-horseradish peroxidase combined with the biotin-labeled ICAM-1 monoclonal antibody, chromogenic substrate 3',3',5,5'-tetramethylbenzidine and ICAM-1 protein standard. A preparation method of the kit comprises the following steps of: (1) preparing the perforated plate coated with the ICAM-1 monoclonal antibody; (2) preparing the biotin-labeled ICAM-1 monoclonal antibody examination fluid; and (3) preparing the aidin-horseradish peroxidase, the chromogenic substrate 3',3',5,5'-tetramethylbenzidine and the ICAM-1 protein standard.

Description

technical field [0001] The invention belongs to the field of medical kits, and in particular relates to a sandwich enzyme-linked immunosorbent assay kit for detecting premature rupture of fetal membranes (PROM) in pregnant women and a preparation method thereof. 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 pregnancy) 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 maternal infection and fetal lung development problems, the...

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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