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Diagnostic device for distingushing between normal and ectopic pregnancy and method for preparing the same

A technology for pregnancy diagnosis and ectopic pregnancy, which is applied in the direction of diagnosis, disease diagnosis, and measuring devices, and can solve the problems of difficult access and effective early identification of pregnant women's risks

Inactive Publication Date: 2003-10-01
HUMASIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even if it is possible to distinguish between normal and abnormal pregnancies, another problem is that effective early differentiation between common spontaneous abortion and ectopic pregnancy, which is dangerous for pregnant women, is difficult to obtain

Method used

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  • Diagnostic device for distingushing between normal and ectopic pregnancy and method for preparing the same
  • Diagnostic device for distingushing between normal and ectopic pregnancy and method for preparing the same
  • Diagnostic device for distingushing between normal and ectopic pregnancy and method for preparing the same

Examples

Experimental program
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Effect test

Embodiment 1

[0031] the scope of the invention. Example 1: Preparation and Purification of Anti-I-hCG Monoclonal Antibody and Anti-α-hCG Monoclonal Antibody A: Immunization and Cell Fusion

[0032] The known cell fusion procedure (Galfre, G et al., 1981, Methods Enzymol. 73:3-46) was used to prepare monoclonal antibodies against I-hCG. First, 20 μg / 100 μl of I-hCG (Zymed, USA) was well emulsified with 100 μl of complete Freund's adjuvant and injected intraperitoneally into Balb / C mice (8 weeks). After 3 weeks, a second intraperitoneal injection was performed under the same protocol as the first injection, except for emulsification with 100 μl of Freund's incomplete adjuvant. After 1 week, blood samples were collected from the mice and antibody formation was determined by ELISA, after which 20 μg hCG was injected into the tail vein. After 3 days, splenocytes were recovered from the mice and subjected to cell fusion with pre-cultured Sp2 / O cells and PEG. Confluent cells were cultured in 9...

Embodiment 2

[0034] Splenocytes were obtained from mice that had previously been immunized three times with β-hCG antigen purchased from Zymed according to the same procedure as in Example 1, A. These cells were mixed with Sp2 / O cells and PEG was used for cell fusion. Confluent cells were cultured in 96-well plates supplemented with HAT medium. Next, cells secreting antibodies reactive to β-hCG and cells secreting antibodies reactive to modified hCGs but not to intact hCG were selected and cultured on a large scale. Next, the monoclonal antibody was purified from the large-scale culture according to the same purification procedure as in Example 1, B, dialyzed with phosphate buffer, adjusted for concentration, and then used for the preparation of a diagnostic kit. Embodiment 3: the preparation of colloidal gold (colored particle)

Embodiment 3

[0035]Colloidal gold with a particle diameter of 20 to 60 nm is used as the colored particles. To prepare colloidal gold, add 220 ml of double distilled water to a 500 ml round bottom flask. The flask was then placed on a hot plate (Corning, USA) and fitted with a reflux device (Pyrex, USA) to prevent evaporation of water. Turn on the hot plate to heat the suspended flask to 100 °C. When the temperature of the distilled water exceeded 100°C, 1.0 ml of 2% gold chloride (Sigma, USA) was added with mixing, followed by 2.0 ml of 1% sodium citrate (Sigma, USA). Continue heating for 30 minutes to produce colloidal gold. The colloidal gold thus obtained was filtered on 0.45 μm filter paper to remove impurities and aggregates, and then used in the preparation of diagnostic devices. Example 4: Preparation of Pads of Colored Particles Incorporating Colored Particles to which Monoclonal Antibodies are Binded

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Abstract

The present invention provides a one-step pregnancy diagnostic device capable of simultaneously detecting and distinguishing normal pregnancy and ectopic pregnancy and a method for preparing such a device. According to the present invention, normal pregnancy and ectopic pregnancy can be quickly and accurately determined at an early stage by immunologically detecting the morphological difference between human chorionic gonadotropin (hCG) and its modified forms secreted into the body fluids of pregnant women.

Description

technical field [0001] The present invention relates to a one-step pregnancy diagnostic device for differentiating normal pregnancy from ectopic pregnancy and a method for the preparation of such a device. More specifically, the present invention relates to the rapid and accurate determination of normal pregnancy and ectopic pregnancy at an early stage by immunodetection of morphological differences between human chorionic gonadotropin (hCG) and its modified forms secreted into the body fluids of pregnant women. Diagnostic devices, and methods of making such devices. Background technique [0002] Ectopic pregnancy refers to the pregnancy in which the fertilized egg implants outside the normal endometrium. According to the position of implantation, it is divided into tubal pregnancy, cervical pregnancy, ovarian pregnancy, peritoneal pregnancy and so on. More than 95% of ectopic pregnancy events are tubal pregnancy, so it is used as a general name for ectopic pregnancy. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53A61B5/00A61B8/08A61B10/00G01N33/50G01N33/531G01N33/543G01N33/558G01N33/577G01N33/68G01N33/76
CPCA61B8/0866A61B10/0012G01N33/558G01N33/689G01N33/76G01N2800/368Y10S436/818A61B5/00G01N33/54388
Inventor 张震东车政学南正铉
Owner HUMASIS
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