CN106124489A

CN106124489A - One can automation mechanized operation, repeatability preferably rheumatoid factor detection reagent box

Info

Publication number: CN106124489A
Application number: CN201610737922.XA
Authority: CN
Inventors: 甘宜梧; 罗维晓; 谭柏清; 王进; 李静
Original assignee: Biobase Biodustry Shandong Co Ltd
Current assignee: Biobase Biodustry Shandong Co Ltd
Priority date: 2016-08-27
Filing date: 2016-08-27
Publication date: 2016-11-16

Abstract

One can automation mechanized operation, repeatability preferably rheumatoid factor detection reagent box.The present invention relates to rheumatoid factor detection technique field, particularly to a kind of rheumatoid factor detection reagent, containing buffer in reagent R1, sodium chloride, Liquid BPF aN3；Containing buffer in reagent R2, glycine, mannitol, trehalose, latex, preservative；Using phosphate buffer, be self-control and domestic production owing to preparing the most of source chemicals used by this test kit, therefore its production cost is low, and the cost of test kit is less than import reagent 80%.

Description

One can automation mechanized operation, repeatability preferably rheumatoid factor detection reagent box

Technical field

The present invention relates to rheumatoid factor detection technique field, particularly to a kind of rheumatoid factor detection reagent, including Use the detection method of this detectable.

Background technology

Rheumatoid arthritis is a kind of common rheumatism, and prevalence is about 0.4%-1.0%, this disease as can not and Time diagnosis and treatment, there is joint deformity, has a strong impact on routine work and life in Chang Qi 1-3 after being ill, improves it and examines in early days Disconnected rate is the problem that medical personnel is paid close attention to；Rheumatoid factor (rheumatoid factor, RF) is that diagnostics classes rheumatic is closed One of major criterion that joint is scorching, diagnosis and prevention are had very important significance by the detection of quickly, efficiently and accurately ground.

Current detection method has: Latex Agglutination, ELISA method and immunoturbidimetry (turbidimetry, scattering turbidimetry Method, latex-enhanced turbidimetry).Latex Agglutination is the method measuring RF earlier, but is only capable of RF is carried out semiquantitative determination, and IgM-RF can only be measured；RF can be quantitative determined by latter two method, and ELISA method can measure RF and hypotype thereof；At present, along with Popularizing of automatic clinical chemistry analyzer, scattered light urbidmetry and turbidimetry are to use more RF assay method in recent years, Establish again the higher latex of sensitivity on the basis of both detection methods and strengthen turbidimetry, but current detectable is external Import, somewhat expensive, develop domestic detection kit the most of fine quality and have great importance for alleviating patient burden.

In consideration of it, it is contemplated that according to the ultimate principle of latex turbidimetry, exploitation has the domestic RF of self-knowledge property right Immue quantitative detection reagent box, the reagent cheap for clinical diagnosis offer, diagnostic value is high.

Summary of the invention

It is an object of the invention to provide a kind of reagent for detecting rheumatoid factor (RF) and use this reagent to detect RF The method of content, the technical specification such as this test kit sensitivity, specificity, stability has all reached to be applied to wanting of Clinical detection Ask, simultaneously because prepare the most of source chemicals used by this test kit to be self-control and domestic production, therefore its production cost Low, the cost of test kit is less than import reagent 80%.

The present invention is obtained through the following steps:

A kind of RF inspection acid test agent, including reagent R1 and reagent R2, the composition of described reagent R1 and reagent R2 is as follows:

1) component of reagent R1 is:

Buffer_{··········································}20mmol/L

Sodium chloride_{··········································}0.1mol/L

Preservative_{··········································}0.5g/L；

2) component of reagent R2 is:

Glycine_{··········································}0.12mol/L

Mannitol_{··········································}10%

Trehalose_{·········································}5%

Latex_{············································}3%

Above reagent is all by the phosphate buffered solution of pH7.5.

Above RF detectable, described preservative is NaN₃。

Described RF detectable detects the detection method of RF, uses automatic clinical chemistry analyzer to utilize performance rate method to carry out Measuring, detection dominant wavelength is 500nm.

Described detection method, the ratio of R1 reagent and R2 reagent is 12:5.

Beneficial effects of the present invention:

1) use new buffer system and stabilizer, significantly improve the stability of reagent；

2) major part source chemicals is self-control and domestic production, and therefore its production cost is low, and the cost of test kit is less than import Reagent 80%；

3) accuracy of reagent and having good stability, easy to use, can meet clinical needs completely；

Figure of description

Fig. 1 embodiment 1 reagent test method

The serum repeatability of the high, normal, basic three kinds of levels of Fig. 2 analyzes (n=20 in crowd)

Fig. 3 serology repeatability analyzes (n=20 between Pi)

The kit results in 4 DEG C of refrigerators of Fig. 4 and 37 DEG C of water bath preservation skies compares

Fig. 5 makes RF test kit and contrast agents box dependency graph by oneself

Fig. 6 makes RF test kit controlled trial by oneself

Fig. 7 embodiment 1 testing result compares with import RF test kit testing result

Detailed description of the invention

Below in conjunction with specific embodiment, the present invention is further described:

Embodiment 1

The detectable of rheumatoid factor, bag reagent R1 and reagent R2:

1) component of reagent R1 is:

2) component of reagent R2 is:

3) using method of the present embodiment reagent:

The RF detectable that the present embodiment describes, uses the automatic clinical chemistry analyzer with double reagent function in use, as Hitachi 7180 fully-automatic analyzers etc., utilize performance rate method to be measured, and according to the ratio of 12:5, R1 and R2 is placed into correspondence On reagent position, the correspondence position at specimen disc places distilled water, standard substance and sample, and operation is such as Fig. 1:

Embodiment 2

Interference is tested:

(1) interference of haemolysis: take the specimen of non-haemolysis, measure RF 3 times respectively and take average, then making it molten with physical method Blood (Hb 500mg/dl), centrifuging and taking supernatant is surveyed RF the most respectively and is taken average 3 times;

(2) interference of blood fat: measure RF tri-times by chylemia specimen (TG 4000mg/dl) the most respectively and take average, being subsequently adding Isopyknic RF calibration solution, measures and takes average for three times and carry out calculating and compare；

(3) interference of yellow cellulitis: first measure the calibration solution of RF each three times and take average, then be separately added into isopyknic concentration and be The Cholerythrin standard liquid of 80mg/dl, measures three times respectively and takes average；

Result display Hb is that the haemolysis specimen of 500mg/dl is noiseless to RF measurement result；When TG is 2000mg/dl without interference with RF measurement result；Concentration is that 40mg/dl bilirubin is to substantially interfering with that RF result mensuration does not has；Blood fat as seen from naked eyes, dilution Measuring RF, result is multiplied by extension rate.

Embodiment 3

RF detection kit parametric measurement

(1) repeated experiment: take the serum sample three of low middle high three kinds of concentration RF, each Concentration Testing 20 times, calculate and criticize Between and variation within batch coefficient (CV%), see Fig. 2, Fig. 3.

Result shows, the repeatability with batch is good, and the reagent of import compares batch interior repeatability zero difference, the repeatability between batch Import reagent is the best, and the self-control test kit of two batches is criticized interior CV and is below 10%, and between batch, CV is less than 15%, meets the biological system of country The regulation (CV < 10%, CV < 20% between crowd in crowd) of product vertification regulation, clinic may be used for this test kit detection RF.

(2) stability test: 4 DEG C of refrigerators will be placed respectively with batch of test kit and 37 DEG C of water baths preserve 10 days laggard The contrast of row RF detection, CV value is more than 10%, assert that reagent lost efficacy.

Test kit preserves 10 days 37 DEG C of water-baths, with 4 DEG C of Refrigerator store kit measurement RF effects without significant change (P < 0.01), illustrate can stably preserve a more than December at 4 DEG C of refrigerators, see Fig. 4.

Embodiment 4

Dependency is tested: utilizes embodiment 1 formula reagent preparation, carries out control test with import RF test kit, have detected simultaneously 40 clinical serum samples, testing result is as shown in Fig. 5, Fig. 6, Fig. 7, and obtains the correlation curve of two kinds of reagent, Y= 1.01X+0.3027, is shown by testing result, and the correlation coefficient of two test kits is 0.9983, illustrates that both have greatly Dependency.

Claims

1. a rheumatoid factor detection reagent, it is characterised in that include reagent R1 and reagent R2, described reagent R1 and reagent R2 Composition as follows:

1) component of reagent R1 is:

Buffer_{···············································}20mol/L

Sodium chloride_{···············································}0.1mol/L

Preservative_{···············································}0.5g/L；

2) component of reagent R2 is:

Glycine_{···············································}0.12mol/L

Mannitol_{···············································}10%

Trehalose_{··············································}5%

Latex_{·················································}3%

Preservative_{···············································}0.5g/L。

Rheumatoid factor detection reagent the most according to claim 1, it is characterised in that in reagent R1, buffer is 25 DEG C, pH It it is the phosphate buffer of 7.5.

Rheumatoid factor detection reagent the most according to claim 1, it is characterised in that in reagent R2, buffer is 25 DEG C, pH It it is the phosphate buffer of 7.5.

Rheumatoid factor detection reagent the most according to claim 1, it is characterised in that described preservative is NaN₃。

5. the rheumatoid factor detection reagent used according to any one of claim 1-4 is to detect the inspection of rheumatoid factor Survey method, it is characterised in that using automatic clinical chemistry analyzer to utilize performance rate method to be measured, detection dominant wavelength is 500nm.

Detection method the most according to claim 5, it is characterised in that the ratio of R1 reagent and R2 reagent is 12:5.