CN202854147U - Pepsinogen II time-resolved fluoresence immunoassay kit - Google Patents

Abstract

The utility model discloses a pepsinogen II time-resolved fluoresence immunoassay kit belonging to the technical field of in-vitro reagent diagnosis. The pepsinogen II time-resolved fluoresence immunoassay kit disclosed by the utility model is based on a double-antibody sandwich method, and antigen-antibody specific immunoreaction is utilized for quantitatively detecting the content of PGII (pepsinogen II) in serum (plasma). An anti-PGII monoclonal antibody-coated plate, a PGII reference standard product, an Eu standard antibody PGII monoclonal antibody, a buffer solution, an enhancement solution and a concentrated plate washing solution are included in the kit. The fluorescence of a marker and the monoclonal antibody with high titer are utilized, so that the purposes of performing direct reaction detection without diluting the serum and reducing workload can be achieved; and simultaneously, a wide measurement range is obtained for facilitating clinical use. The pepsinogen II time-resolved fluoresence immunoassay kit disclosed by the utility model has the positive significances of high sensitivity, good specificity, wide measurement range, simplicity in operation and broad market prospects, and is conductive to large-scale popularization and application.

Description

A kind of pepsinogen I I time-resolved fluoroimmunoassay detection kit

Technical field

The utility model relates to time resolved fluoro-immunoassay (TRFIA) kit of pepsinogen I I content in the quantitative detection serum (blood plasma), belongs to external reagent diagnosis technical field.

Background technology

PGⅡ (being called for short the PG II) derives from full gastric gland and distal duodenum BrunnerShi gland, the synthetic PGII of gastric mucosa is about 25% of total amount, PG II major part enters alimentary canal, enters on a small quantity blood circulation, therefore is called as " pointer of reflection stomach state and function ".

Clinical research both at home and abroad points out that the correlativity of PG II and gastric mucosa pathology is larger, and its rising and fundus gland shrink tube, intestinal metaplasia or false gland metaplasia, dysplasia are relevant.Measure PG II content and help other disease of digestive tracts such as detecting duodenal ulcer, atrophic gastritis, gastritis, cancer of the stomach, for clinical detection and health check-up examination demand.In the examination of crowd's stomach trouble, everyone makees gastroscope is unpractical, can detect with high-risk Mass screenings such as superficial gastritis, erosive gastritis, gastric ulcer, duodenal ulcer, cancer of the stomach out by the Noninvasive serum PG, carrying out gastrocopy is a kind of realistic plan again.

Adopt the time-resolved fluorescence measuring technique, take rare earth ion as tracer material, the TRFIIA kit of the PGII of setting up has measures highly sensitive, easy and simple to handle, the advantages such as tracer stable, quantitative test broad quantum, no radioactivity pollute, be one of the sensitiveest in the present PGII detection method, that measurement range is the widest method, be very beneficial for large-scale generaI investigation screening and patient's course of disease and detect.

The utility model content

The purpose of this utility model provides the time-resolved fluoroimmunoassay detection kit of a kind of novel PGII, uses the Fluorescence amplification effect of rare earth ion,, reach purpose highly sensitive, that specificity good, range ability is wide.This kit can be realized quick, easy detection.

The technical solution of the utility model: a kind of PGⅡ time-resolved fluoroimmunoassay detection kit, the box body of this kit is provided with in (7): be coated with the microwell plate of PG II monoclonal antibody, i.e. 1 of antibody response plate (1); 6 bottles of PG II standard items (2); Eu marks anti-PG II monoclonal antibody (3), damping fluid 1 bottle of (4), enhancing liquid 1 bottle of (5), concentrated 1 bottle of the plate liquid (6) of washing;

Antibody response plate 1 (1) is positioned over the right front portion in the box body (7), and PG II standard items 6 bottles (2) are positioned over the right back section in the box body (7), are the bottle of 6 placed adjacent of placing vertically; Place 1 Eu that places vertically and mark anti-PG II monoclonal antibody (3) bottle for left back in the box body (7), left front section in the box body (7) places horizontal damping fluid 1 bottle of (4), concentrated plate liquid 1 bottle of (6), enhancing 1 bottle of liquid (5) washed from front to back successively, this kind modes of emplacement makes the kit volume compact, and easy to use;

Antibody response plate (1) is that anti-PG II monoclonal antibody is coated, detachable 96 orifice plates; Eu marks anti-PG II monoclonal antibody (3) and is the paired anti-PG II monoclonal antibody of another strain of Eu mark and coated antibody; PG II standard items (2) are PG II antigen dried frozen aquatic products one cover of variable concentrations.

Attached operational manual in addition.The reagent of each kit is measured for enough 96 times, antigen during detection in coated antibody and another strain antibody of Eu mark and the sample consists of sandwich complex, and after adding strengthened liquid, the fluorescent counting became positive correlation with sample P GII content, namely obtain corresponding PGII concentration according to typical curve, easy and simple to handle.

The beneficial effects of the utility model: one, highly sensitive, can reach 0.02ng/mL; Two, broad quantum, serum (slurry) sample do not need dilution, and the sample concentration value can both accurately detect between 0-50ng/mL's; Three, lack detection time, finished in 2 hours.

Description of drawings

Fig. 1 structural representation of the present utility model.Among the figure: 1, be coated with the microwell plate of PGII monoclonal antibody, i.e. 1 of antibody response plate (96T); 2, PGII standard items (0-50ng/mL, 6 bottles); 3, Eu marks anti-PGII monoclonal antibody (another strain, paired with coated antibody) 1 * 0.5mL/ bottle (time spent 1:50 dilution); 4, damping fluid: one bottle, 50mL; 5, strengthen liquid: one bottle, 30mL; 6, the concentrated plate liquid of washing: one bottle, 40mL; 7, box body.

Embodiment

Embodiment 1:PGII TRFIA sample determination

(1) preparation of reagent

A, dried frozen aquatic products: before using with the dried frozen aquatic products balance to room temperature, use the 1mL dissolved in distilled water.

B, use forward horizontal stand to room temperature the antibody response lath of requirement.

C, cleansing solution: will concentrate and wash plate liquid with 25 times of dilutions of distilled water 1 ︰, as the work cleansing solution.

D, Eu-mark the dilution of anti-PGII monoclonal antibody: measure per sample 50 times of dilutions with damping fluid 1 ︰ in application in front 1 hour.

(2) test procedure

A, get the antibody response plate, since the first hole, every hole adds the normative reference product (totally 6 standard points, suggestion is done multiple hole to guarantee the reliability of typical curve) of 50 μ L successively.Behind the normative reference product, add successively 50 μ L samples (suggestion adopts diplopore to measure).

C, add after normative reference product and the sample, every hole adds 150 μ L damping fluids again, 25 ℃ of oscillation incubations 1 hour, work cleansing solution flushing 4 times.

The Eu that d, every hole add successively with 50 times of dilutions of damping fluid 1 ︰ marks anti-PGII monoclonal antibody 200 μ L, 25 ℃ of oscillation incubations 1 hour, work cleansing solution flushing 6 times.

E, every hole add enhancing liquid 200 μ L, and 25 ℃ of oscillation incubations were measured after 5 minutes.

Behind f, the end of run to be tested, withdraw from sample, regain agents useful for same in 2 ~ 8 ℃ of placements.Used antibody response lath discards after taking out, and please don't reuse.

Result treatment: the corresponding luminous intensity according to the serial reference standard items of known PGII concentration can obtain typical curve, return to process to obtain regression straight line by the double-log mathematical model, the PGII concentration of unknown sample can be by its corresponding luminous intensity inverse out (table 1) from the regression straight line.

Table 1

Embodiment described in the utility model is guiding; indefiniteness; just help to understand the utility model; therefore the utility model is not limited to the embodiment described in the embodiment; every by those skilled in the art according to other embodiments that the technical solution of the utility model draws, belong to equally the scope of the utility model protection.

Claims (1)

1. PGⅡ time-resolved fluoroimmunoassay detection kit, it is characterized in that: the box body of kit is provided with in (7): be coated with the microwell plate of PG II monoclonal antibody, i.e. 1 of antibody response plate (1); 6 bottles of PG II standard items (2); Eu marks anti-PG II monoclonal antibody (3), damping fluid 1 bottle of (4), enhancing liquid 1 bottle of (5), concentrated 1 bottle of the plate liquid (6) of washing;

Antibody response plate 1 (1) is positioned over the right front portion in the box body (7), and PG II standard items 6 bottles (2) are positioned over the right back section in the box body (7), are the bottle of 6 placed adjacent of placing vertically; Place 1 Eu that places vertically and mark anti-PG II monoclonal antibody (3) bottle for left back in the box body (7), left front section in the box body (7) places horizontal damping fluid 1 bottle of (4), concentrated plate liquid 1 bottle of (6), enhancing 1 bottle of liquid (5) washed from front to back successively, this kind modes of emplacement makes the kit volume compact, and easy to use;

Antibody response plate (1) is that anti-PG II monoclonal antibody is coated, detachable 96 orifice plates; Eu marks anti-PG II monoclonal antibody (3) and is the paired anti-PG II monoclonal antibody of another strain of Eu mark and coated antibody; PG II standard items (2) are PG II antigen dried frozen aquatic products one cover of variable concentrations.

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