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

An ICAM-1 and detection kit technology, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems such as the preparation method of the detection kit for premature rupture of membranes that has not yet been found, and achieve the effect of improving the accuracy

Active Publication Date: 2010-10-27
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
  • 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 a premature rupture of membranes detection kit with ICAM-1 as the detection index:

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

[0047] Dilute the ICAM-1 monoclonal antibody with a carbonate buffer of pH=9 to a dilution with a concentration of 10μg / ml, and then add the ICAM-1 monoclonal antibody at a volume of 100μl / well to the 96-well plate In each well, coat at 4°C for 12 hours; after the coating time expires, add 120μl / well of bovine serum albumin solution with a concentration of 1g / 100ml to each well on the multiwell plate and perform a blocking reaction at 37°C. The reaction time is 1 hour; after the blocking reaction, wash the multiwell plate with pH=7.2 and a concentration of 0.05 mol / l phosphate buffer. When the unreacted substances on the multiwell plate are removed, the coated ICAM-1 is obtained. The multi-well plate of monoclonal antibody, then put it in t...

Embodiment 2

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

[0054] 1. Samples to be tested

[0055] (1) Standard sample preparation

[0056] Set the ICAM-1 protein standard prepared in the kit described in Example 1 as the standard with the highest concentration of ICAM-1, named as Standard 1, the concentration of ICAM-1 is 10ng / ml, and then use the example 1 The sample diluent provided in the kit dilutes the ICAM-1 protein standard product into ICAM-1 concentrations of 5ng / ml, 2.5ng / ml, 1.25ng / ml, 0.63ng / ml, 0.32ng / The standard samples of ml and 0.16ng / ml are named Standard Sample 2, Standard Sample 3, Standard Sample 4, Standard Sample 5, Standard Sample 6, and Standard Sample 7, a total of 7 standard samples.

[0057] (2) Sample

[0058] Taking parturients from the outpatient department and hospitalization of West China Second Hospital of Sichuan University as the experimental subjects, the subjects were divi...

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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 Membrane (PROM) in pregnant women and a preparation method thereof. Background technique [0002] Rupture of Fetal Membrane (ROM) may occur at any time during pregnancy. Rupture of Fetal Membrane that occurs before labor is called Premature Rupture of Fetal Membrane (PROM). The incidence of PROM after term (37 gestational weeks) is about 10%; the incidence of PROM before term (37 gestational weeks) is 2-3.5%. Premature rupture of membranes is the main factor leading to prenatal and postpartum complications in obstetric patients, and the main reason for premature delivery of fetuses and neonatal admission to intensive care unit. Since medical staff have to balance the prolonged gestational week with the risk of intrauterine and pregnant women’s infections and fetal lung development problem...

Claims

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Application Information

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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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