CN103196727A - Blood zinc protoporphyrin whole blood quality control product and preparation method and applications thereof - Google Patents
Blood zinc protoporphyrin whole blood quality control product and preparation method and applications thereof Download PDFInfo
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- CN103196727A CN103196727A CN2013101206558A CN201310120655A CN103196727A CN 103196727 A CN103196727 A CN 103196727A CN 2013101206558 A CN2013101206558 A CN 2013101206558A CN 201310120655 A CN201310120655 A CN 201310120655A CN 103196727 A CN103196727 A CN 103196727A
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Abstract
The invention discloses a blood zinc protoporphyrin whole blood quality control product and a preparation method and applications thereof. For the quality control product, the anti-freezing whole blood of an invitro non-lead poisoning average people or lead poisoning patients is adopted as solvent, wherein the final concentration of sodium azide is 0.15-0.25mu g/mL. The anti-freezing whole blood is whole blood in which the final concentration of the EDTA-2K {(ethylenedinitrilo)tetraacetate dipotassium salt} IS 1mg/mL. The quality control product can simultaneously detect the normal level and high-level sample detection precision of instruments. The blood zinc protoporphyrin whole blood quality control product can remove matrix effect, is easy for source, simple and convenient to prepare, fewer in additives, low in cost, good in stability during storage period, convenient to prepare and use in a short time in a laboratory, and applicable to monitoring on the accuracy and precision of a blood zinc protoporphyrin determinator.
Description
Technical field
The invention belongs to biological product technical field, be specifically related to a kind of blood zinc protoporphyrin whole blood quality control product and preparation method thereof and application.
Background technology
In recent years, the occupational health monitoring more and more comes into one's own, and saturnism is a big difficult point of occupational health monitoring, and the main foundation of occupational chronic saturnism diagnostic criteria is that index is detected in blood lead and blood zinc protoporphyrin laboratories such as (ZPP).The instrument and equipment condition that the blood lead detection needs can not be used for large tracts of land examination and popularization than higher; Fluorometric assay technology for detection blood zinc protoporphyrin has advantages such as quick, accurate, inexpensive, has been widely used in hypoferric anemia and saturnism diagnosis.Studies show that blood zinc protoporphyrin and blood lead and lead in urine do not have difference as the plumbous harm of occupation monitoring index, therefore, the blood zinc protoporphyrin detects the great attention that is subjected to occupational hazards appraisal person.
Accurately accurate occupational illness laboratory detection data are one of main foundations of diagnosis of occupational disease evaluation, and its acquisition needs the indoor Quality Control through strictness.Indoor Quality Control is that the staff by the laboratory adopts a series of statistical methods, estimates the degree of reliability of this experimental determination work continuously, judges the process whether survey report can send.The purpose of indoor Quality Control is to detect, control the precision of this experimental determination work, and detects the change of its accuracy, improve conventional determining work batch between, batch in the consistance of detection result of specimen.Quality-control product is to be specifically designed to the material approximate with sample to be measured indoor Quality Control, stable, is the important substance basis that guarantees Quality Control work.
At present, the blood zinc protoporphyrin detects does not still have commercial indoor quality-control product, and it is very necessary to develop the indoor quality-control product of a kind of blood zinc protoporphyrin.
Summary of the invention
The purpose of this invention is to provide a kind of blood zinc protoporphyrin whole blood quality control product, described quality-control product be the anticoagulated whole blood that contains zinc protoporphyrin be solvent, wherein the final concentration of Sodium azide is 0.15-0.25 μ g/mL, as 0.15-0.20 μ g/mL or 0.20-0.25 μ g/mL.
In above-mentioned quality-control product, described anticoagulated whole blood can be the EDTA-2K(EDTAP dipotassium ethylene diamine tetraacetate) final concentration is the whole blood of 1mg/mL.
In above-mentioned quality-control product, the described anticoagulated whole blood that contains zinc protoporphyrin is not for to add the anticoagulated whole blood of zinc protoporphyrin, specifically can be stripped normal person's (non-saturnism) anticoagulated whole blood or stripped saturnism patient's anticoagulated whole blood.
The present invention also provides a kind of preparation method of blood zinc protoporphyrin whole blood quality control product, comprises that the whole blood that will contain zinc protoporphyrin carries out anti-freezing and handles, and obtains anticoagulated whole blood; Again by the volume ratio of 500:1 add in the described anticoagulated whole blood concentration be 0.075-0.125g/L(as the sodium azide aqueous solution of 0.075-0.1g/L or 0.1-0.125g/L), obtain described blood zinc protoporphyrin whole blood quality control product.
In said method, it is according to the ratio of 10mL:10mg the described whole blood that contains zinc protoporphyrin to be mixed with EDTA-2K that described anti-freezing is handled.
In said method, the described anticoagulated whole blood that contains zinc protoporphyrin is not for to add the anticoagulated whole blood of zinc protoporphyrin, specifically can be stripped normal person's (non-saturnism) whole blood or stripped saturnism patient's whole blood.
The present invention protects the application of arbitrary described blood zinc protoporphyrin whole blood quality control product in the quantitative detection kit of preparation blood zinc protoporphyrin in the said goods and the method.Described quantitative detection method specifically can be the surface fluorescence method.
The principle that described surface fluorescence standard measure detects the blood zinc protoporphyrin is as follows:
Zinc protoporphyrin has the characteristic fluorescence spectrum, namely under the exciting of 425nm incident light, the zinc protoporphyrin emission wavelength is the fluorescence of 594nm, and the content of zinc protoporphyrin is the certain proportion relation in this fluorescence intensity and the testing sample, by measuring this fluorescence intensity, can extrapolate the content of zinc protoporphyrin in the testing sample.
The present invention is main material with the lower normal person's of the content of the higher saturnism patient whole blood of the content of zinc protoporphyrin and zinc protoporphyrin whole blood respectively, after carrying out anti-freezing and handle with the EDTA-2K of 1mg/mL, the Sodium azide that adds 0.15-0.25 μ g/mL again prepares the quality-control product of two levels, it was preserved 30 days down at 2-8 ℃, detect the content of zinc protoporphyrin through the surface fluorescence method, the coefficient of variation can satisfy the general requirement of laboratory precision fully all less than 5%, according to Quality Control rule 1 commonly used
3SWith 2
2SJudge, out-of-control phenomenon do not occur in 30 days.Illustrate and add 0.15-0.25 μ g/mL Sodium azide in the whole blood, can effectively suppress the destruction that bacterium is caused red blood cell.
Quality-control product provided by the present invention monitoring instrument is simultaneously examined the precision of the gentle high-level pattern detection of normal water.Quality-control product provided by the present invention can be got rid of matrix effect, originate easy, simple for production, additive is few, cost is low, stability is fine between storage life, be convenient in a short time prepared in laboratory and use, be applicable to accuracy and the precision monitoring of blood zinc protoporphyrin analyzer.
Description of drawings
Fig. 1 is observations stabilization time of normal value group blood zinc protoporphyrin whole blood quality control product.Wherein, horizontal ordinate is for detecting the date, and ordinate is testing result (unit: μ mol/L).
Fig. 2 is observations stabilization time of high value group blood zinc protoporphyrin whole blood quality control product.Wherein, horizontal ordinate is for detecting the date, and ordinate is testing result (unit: μ mol/L).
Embodiment
Employed experimental technique is conventional method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The employed instrument of zinc protoporphyrin assay among the following embodiment is the ZPP3800 type blood zinc protoporphyrin analyzer of Kangda Development Co., Guangdong Prov., and its assay method is the surface fluorescence method.
Before the quality-control product of 2-8 ℃ of preservations is carrying out the zinc protoporphyrin assay, place 30 minutes balances earlier to room temperature and abundant mixing among the following embodiment.
Related experiment blood is residue blood and ratify through occupational disease precaution clinic, the Shenzhen ethics council behind the blood routine examination among the following embodiment.
Preparation and the estimation of stability of embodiment 1, blood zinc protoporphyrin whole blood quality control product
One, the preparation of blood zinc protoporphyrin whole blood quality control product
1, the preparation of high value group blood zinc protoporphyrin whole blood quality control product
Gather the chronic lead poisoning patient's (result who detects through blood lead is 5.67 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the test tube that contains 10.0mg EDTA-2K, fully behind the mixing, the concentration that adds 20 μ L is the sodium azide aqueous solution of 0.1g/L, 2-8 ℃ of preservations after the packing obtain high value group blood zinc protoporphyrin whole blood quality control product.
2, the preparation of normal value group blood zinc protoporphyrin whole blood quality control product
Gather normal person's's (result who detects through blood lead is 0.32 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, the concentration that adds 20 μ L is the sodium azide aqueous solution of 0.1g/L, 2-8 ℃ of preservations after the packing obtain normal value group blood zinc protoporphyrin whole blood quality control product.
3, the preparation of contrast whole blood quality control product
(result who detects through blood lead is 0.32 μ mol/L to gather the normal person of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index, with step 2 be same people) whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, not packing, direct 2-8 ℃ of preservations obtain contrasting the whole blood quality control product.
Two, detect Sodium azide to the influence of blood zinc protoporphyrin content in the quality-control product
Step 1 is prepared simultaneously, 2-8 ℃ preserve 1 day normal value group blood zinc protoporphyrin whole blood quality control product (L) and contrast whole blood quality control product (CK) and respectively take out 1, every quality-control product serial sampling 10 times, zinc protoporphyrin content in the difference working sample, the result is as shown in table 1.The correlativity of CK and two groups of data of L draws correlation coefficient r=0.955 in the analytical table 1.The result shows that the 0.1g/L Sodium azide that adds in the blood zinc protoporphyrin whole blood quality control product does not have influence to the blood zinc protoporphyrin content in the whole blood.
The blood zinc protoporphyrin content detection result (unit: μ mol/L) of table 1, CK and L
| Number of times | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| CK | 0.90 | 0.90 | 0.85 | 0.92 | 0.92 | 0.90 | 0.92 | 0.90 | 0.90 | 0.94 |
| L | 0.92 | 0.91 | 0.86 | 0.93 | 0.94 | 0.91 | 0.95 | 0.91 | 0.90 | 0.95 |
Three, the estimation of stability of blood zinc protoporphyrin whole blood quality control product
(1) estimation of stability of normal value group blood zinc protoporphyrin whole blood quality control product
1, homogeneity is measured
Get step 1 prepares, 2-8 ℃ of normal value group blood zinc protoporphyrin whole blood quality control product of same batch that preservation is preceding, randomly draw 20, on the same day the zinc protoporphyrin content in each sample of METHOD FOR CONTINUOUS DETERMINATION.Be the quality-control product with homogeneity with the coefficient of variation less than 5% quality-control product candidate.
2, the stability of quality-control product is measured
The quality-control product that step 1 is had homogeneity carries out 20 days continuous detecting (namely preserving 1-20 days through 2-8 ℃), randomly draws a sample every day, measures the zinc protoporphyrin content in each sample, and the result is as shown in table 2.In the table 2 mean value of 20 data be 0.91, standard deviation be 0.04 and the coefficient of variation be 4.32%.
The Detection of Stability result of table 2, normal value group (unit: μ mol/L)
| My god | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| The result | 0.93 | 0.87 | 0.94 | 0.89 | 0.97 | 0.91 | 0.88 | 0.84 | 0.95 | 0.92 |
| My god | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 |
| The result | 0.85 | 0.96 | 0.91 | 0.92 | 0.93 | 0.97 | 0.85 | 0.96 | 0.91 | 0.92 |
3, observe the stabilization time of quality-control product
To measure the quality-control product with homogeneity through step 1 and carry out continuous detecting (namely preserving 1-20 days through 2-8 ℃), randomly draw a sample every day, measure the zinc protoporphyrin content in each sample, calculating mean value, standard deviation and the coefficient of variation, and centered by this mean value line, with mean value ± 2 times standard deviation serve as the warning line, be line out of control with mean value ± 3 times standard deviation, draw the Quality Control frame diagram, the result that later on each (once a day namely) detects clicks and enters in this Quality Control frame diagram, until there being the result who exceeds line out of control to occur, stop experiment, record stabilization time (namely preparing to the fate that toe-in fruit out of control do not occur exceeding from quality-control product).
The result: according to preceding 20 days (the Quality Control frame diagram (Fig. 1) that the result of 12-13～1-1) draws, its mean value is 0.91, standard deviation is 0.03; Through observing, be 34 days the stabilization time of normal value group blood zinc protoporphyrin whole blood quality control product, and the testing result of every day is as shown in table 3.
Observations stabilization time of table 3, normal value group (unit: μ mol/L)
| Date | The result | Date | The result | Date | The result | Date | The result | Date | The result |
| 12-13 | 0.92 | 12-20 | 0.86 | 12-27 | 0.94 | 1-3 | 0.92 | 1-10 | 0.88 |
| 12-14 | 0.86 | 12-21 | 0.92 | 12-28 | 0.92 | 1-4 | 0.95 | 1-11 | 0.93 |
| 12-15 | 0.95 | 12-22 | 0.92 | 12-29 | 0.94 | 1-5 | 0.92 | 1-12 | 0.89 |
| 12-16 | 0.87 | 12-23 | 0.90 | 12-30 | 0.85 | 1-6 | 0.94 | 1-13 | 0.91 |
| 12-17 | 0.96 | 12-24 | 0.92 | 12-31 | 0.90 | 1-7 | 0.91 | 1-14 | 0.94 |
| 12-18 | 0.90 | 12-25 | 0.90 | 1-1 | 0.92 | 1-8 | 0.90 | 1-15 | 0.90 |
| 12-19 | 0.90 | 12-26 | 0.90 | 1-2 | 0.96 | 1-9 | 0.91 | ? | ? |
(2) estimation of stability of high value group blood zinc protoporphyrin whole blood quality control product
1, homogeneity is measured
Get step 1 prepares, 2-8 ℃ of high value group blood zinc protoporphyrin whole blood quality control product of same batch that preservation is preceding, randomly draw 20, on the same day the zinc protoporphyrin content in each sample of METHOD FOR CONTINUOUS DETERMINATION.Be the quality-control product with homogeneity with the coefficient of variation less than 5% quality-control product candidate.
2, the stability of quality-control product is measured
The quality-control product that step 1 is had homogeneity carries out 20 days continuous detecting (namely preserving 1-20 days through 2-8 ℃), randomly draws a sample every day, measures the zinc protoporphyrin content in each sample, and the result is as shown in table 4.In the table 4 mean value of 20 data be 12.10, standard deviation be 0.39 and the coefficient of variation be 3.19%.
The Detection of Stability result of table 4, high value group (unit: μ mol/L)
| My god | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| The result | 11.11 | 11.81 | 11.84 | 12.01 | 11.91 | 11.98 | 11.99 | 12.1 | 12.11 | 11.88 |
| My god | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 |
| The result | 12.12 | 12.1 | 12.14 | 13.01 | 12.21 | 12.12 | 12.1 | 12.14 | 13.01 | 12.21 |
3, observe the stabilization time of quality-control product
3 identical in method and the step ().
The result: according to preceding 20 days (the Quality Control frame diagram (Fig. 2) that the result of 12-13～1-1) draws, its mean value is 11.95, standard deviation is 0.50; Through observing, be 33 days the stabilization time of high value group blood zinc protoporphyrin whole blood quality control product, and the testing result of every day is as shown in table 5.
Observations stabilization time of table 5, high value group (unit: μ mol/L)
| Date | The result | Date | The result | Date | The result | Date | The result | Date | The result |
| 12-13 | 11.21 | 12-20 | 12.14 | 12-27 | 12.10 | 1-3 | 11.64 | 1-10 | 11.71 |
| 12-14 | 11.60 | 12-21 | 12.21 | 12-28 | 12.82 | 1-4 | 11.66 | 1-11 | 11.76 |
| 12-15 | 11.94 | 12-22 | 11.80 | 12-29 | 12.21 | 1-5 | 11.59 | 1-12 | 11.96 |
| 12-16 | 12.05 | 12-23 | 12.14 | 12-30 | 11.80 | 1-6 | 11.91 | 1-13 | 11.51 |
| 12-17 | 11.94 | 12-24 | 12.11 | 12-31 | 11.54 | 1-7 | 11.83 | 1-14 | 11.31 |
| 12-18 | 11.96 | 12-25 | 12.13 | 1-1 | 11.54 | 1-8 | 11.85 | 1-15 | 13.88 |
| 12-19 | 11.89 | 12-26 | 13.13 | 1-2 | 11.74 | 1-9 | 11.72 | ? | ? |
Experiment in the step 3 respectively repeats respectively 3 times, and each quality-control product that repeats with same batch carries out, and the stabilization time of each batch is all more than 30 days as a result, and coefficient of variation CV is less than 5%, according to Quality Control rule 1 commonly used
3SWith 2
2SJudge, out-of-control phenomenon all do not occur in 30 days.
Preparation and the estimation of stability of embodiment 2, blood zinc protoporphyrin whole blood quality control product
One, the preparation of blood zinc protoporphyrin whole blood quality control product
1, the preparation of high value group blood zinc protoporphyrin whole blood quality control product
Gather the chronic lead poisoning patient's (result who detects through blood lead is 5.67 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the test tube that contains 10.0mg EDTA-2K, fully behind the mixing, the concentration that adds 20 μ L is
0.075g/L sodium azide aqueous solution, 2-8 ℃ of preservations after the packing obtain high value group blood zinc protoporphyrin whole blood quality control product.
2, the preparation of normal value group blood zinc protoporphyrin whole blood quality control product
Gather normal person's's (result who detects through blood lead is 0.32 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, the concentration that adds 20 μ L is
0.075g/L sodium azide aqueous solution, 2-8 ℃ of preservations after the packing obtain normal value group blood zinc protoporphyrin whole blood quality control product.
3, the preparation of contrast whole blood quality control product
(result who detects through blood lead is 0.32 μ mol/L to gather the normal person of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index, with step 2 be same people) whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, not packing, direct 2-8 ℃ of preservations obtain contrasting the whole blood quality control product.
Two, detect Sodium azide to the influence of blood zinc protoporphyrin content in the quality-control product
Method and result are identical with step 2 among the embodiment 1.
Three, the estimation of stability of blood zinc protoporphyrin whole blood quality control product
Method and result are identical with step 3 among the embodiment 1.
Preparation and the estimation of stability of embodiment 3, blood zinc protoporphyrin whole blood quality control product
One, the preparation of blood zinc protoporphyrin whole blood quality control product
1, the preparation of high value group blood zinc protoporphyrin whole blood quality control product
Gather the chronic lead poisoning patient's (result who detects through blood lead is 5.67 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the test tube that contains 10.0mg EDTA-2K, fully behind the mixing, the concentration that adds 20 μ L is
0.125g/L sodium azide aqueous solution, 2-8 ℃ of preservations after the packing obtain high value group blood zinc protoporphyrin whole blood quality control product.
2, the preparation of normal value group blood zinc protoporphyrin whole blood quality control product
Gather normal person's's (result who detects through blood lead is 0.32 μ mol/L) of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, the concentration that adds 20 μ L is
0.125g/L sodium azide aqueous solution, 2-8 ℃ of preservations after the packing obtain normal value group blood zinc protoporphyrin whole blood quality control product.
3, the preparation of contrast whole blood quality control product
(result who detects through blood lead is 0.32 μ mol/L to gather the normal person of clinical no haemolysis, turbid, the no jaundice of no fat, no bacteria pollution and other infectiousness index, with step 2 be same people) whole blood 10ml, collect with the EDTA-2K test tube that contains 10.0mg, fully behind the mixing, not packing, direct 2-8 ℃ of preservations obtain contrasting the whole blood quality control product.
Two, detect Sodium azide to the influence of blood zinc protoporphyrin content in the quality-control product
Method and result are identical with step 2 among the embodiment 1.
Three, the estimation of stability of blood zinc protoporphyrin whole blood quality control product
Method and result are identical with step 3 among the embodiment 1.
Claims (10)
1. blood zinc protoporphyrin whole blood quality control product is characterized in that: described quality-control product be the anticoagulated whole blood that contains zinc protoporphyrin be solvent, wherein the final concentration of Sodium azide is 0.15-0.25 μ g/mL.
2. quality-control product according to claim 1, it is characterized in that: described anticoagulated whole blood is the whole blood of 1mg/mL for the EDTA-2K final concentration.
3. quality-control product according to claim 1 and 2 is characterized in that: the described anticoagulated whole blood of zinc protoporphyrin that contains is not for adding the anticoagulated whole blood of zinc protoporphyrin.
4. according to arbitrary described quality-control product in the claim 1-3, it is characterized in that: the described anticoagulated whole blood that contains zinc protoporphyrin is the normal person's that exsomatizes anticoagulated whole blood or stripped saturnism patient's anticoagulated whole blood.
5. the preparation method of blood zinc protoporphyrin whole blood quality control product comprises that the whole blood that contains zinc protoporphyrin that will exsomatize carries out anti-freezing and handles, and obtains anticoagulated whole blood; Adding concentration by the volume ratio of 500:1 in the described anticoagulated whole blood again is the sodium azide aqueous solution of 0.075-0.125g/L, obtains described blood zinc protoporphyrin whole blood quality control product.
6. method according to claim 5 is characterized in that: it is according to the ratio of 10mL:10mg the described stripped whole blood that contains zinc protoporphyrin to be mixed with EDTA-2K that described anti-freezing is handled.
7. according to claim 5 or 6 described methods, it is characterized in that: the described whole blood of zinc protoporphyrin that contains is not for adding the whole blood of zinc protoporphyrin.
8. according to arbitrary described method in the claim 4-7, it is characterized in that: the described whole blood that contains zinc protoporphyrin is the normal person's that exsomatizes whole blood or stripped saturnism patient's whole blood.
9. the application of the arbitrary described blood zinc protoporphyrin whole blood quality control product in the claim 1-4 or 5-8 in the quantitative detection kit of preparation blood zinc protoporphyrin.
10. application according to claim 9 is characterized in that: described quantitative detection method is the surface fluorescence method.
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| CN105498000A (en) * | 2016-01-20 | 2016-04-20 | 廊坊市中心血站 | Method-plasma quality control chart for ensuring that blood quality meets blood transfusion effect |
| CN107367409A (en) * | 2017-07-04 | 2017-11-21 | 长沙金域医学检验所有限公司 | The quantitative Internal Quality Control product of whole blood EBV DNA |
| CN110763527A (en) * | 2018-07-25 | 2020-02-07 | 苏州迈瑞科技有限公司 | Urine visible component quality control product and preparation method and application thereof |
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| CN105498000B (en) * | 2016-01-20 | 2017-11-07 | 廊坊市中心血站 | It is a kind of to ensure the method blood plasma Quality Control figure that blood quality meets blood transfusion effect |
| CN107367409A (en) * | 2017-07-04 | 2017-11-21 | 长沙金域医学检验所有限公司 | The quantitative Internal Quality Control product of whole blood EBV DNA |
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