CN1156707C - Children's lead poisoning screening paper sheet method - Google Patents
Children's lead poisoning screening paper sheet method Download PDFInfo
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- CN1156707C CN1156707C CNB001154117A CN00115411A CN1156707C CN 1156707 C CN1156707 C CN 1156707C CN B001154117 A CNB001154117 A CN B001154117A CN 00115411 A CN00115411 A CN 00115411A CN 1156707 C CN1156707 C CN 1156707C
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- blood
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Abstract
The present invention provides a paper sheet method for screening lead poisoning children, which relates to a blood lead detection method. The method comprises blood sample collection, pretreatment, standard curve establishment and tests. The method is characterized in that the method comprises: step one, collecting a blood sample from the finger pulp, forming a blood spot with the diameter larger than 0.6cm on filter paper, airing the filter paper at room temperature, successively putting the aired filter paper in a deionized plastic bag, and preserving the filter paper in the plastic bag at the environmental temperature between 4 DEG C and 10 DEG C; step two, pretreating the sample: holing the center of the blood spot, cutting off a small circular sheet with the diameter of 0.6cm, directly putting the small circular sheet into a test tube, adding 200 ul of nitric acid with 3 to 10% of concentration to the test tube, shaking the test tube for 5 to 10 seconds to submerge the paper sheet fully into the nitrite acid solution, placing the test tube at the environmental temperature of 4 DEG C for 16 to 24 hours, taking out the test tube, successively shaking the test tube for 5 to 10 seconds, then centrifugally processing the solution, and taking supernatant fluid for tests; step three, establishing a standard curve: selecting five to six points within the known blood lead concentration range of 0 to 500 mug/L, dripping 30 ul of the blood sample on filter paper, airing the filter paper at room temperature, successively carrying out sample pretreatment as the test sample is treated in step two, testing and establishing a curve.
Description
Affiliated technical field
The method of paper disk method examination children ' s lead poisoning of the present invention relates to a kind of blood lead detection method.
Background technology
The method of domestic blood lead test all need adopt venous blood or tip blood, and blood specimen must be put into test tube and could transport, and makes this problem of transportation difficulty very outstanding, and the area of no checkout equipment can't be carried out the detection of blood lead substantially.There is paper disk method to carry out blood lead abroad and detects, make this difficult problem of transportation difficulty of sample obtain solution, the foundation that its step is blood sampling, sample pre-service, typical curve, usefulness Atomic Absorption Spectrometer (graphite furnace method) test.Wherein blood collection procedure is that one 1/2 inch circle is arranged on filter paper, gets the 6-8 full circle of bleeding, and this blood collection procedure has blood sampling difficulty, easily overlapping, the blood coagulation of drop of blood, thereby influences test result; The sample preprocessing process is that 2 of the samples of 0.6cm diameter are put in vitro, puts into 5% nitric acid 0.6ml, and vibration was left standstill 15 seconds, repeats to vibrate for 2 times and leave standstill process, and is centrifugal, gets supernatant mensuration.This sample preprocessing process number of oscillation is many, and pretreatment time is long, and it is many to get specimen amount, and revision test is difficulty.The process of setting up of typical curve is that the standard water solution dilution with known lead concentration becomes 0,10,20,50,100 μ g/dl, add 2 on the blank scraps of paper, make the usefulness of typical curve during for test, make typical curve with aqueous solution, inconsistent with the blood specimen background of test, error is bigger; In test process, application of sample 16 μ l add biphosphate sodium 7 μ l, and sample size is big, and the test duration is long.Simultaneously, the Atomic Absorption Spectrometer that is used for blood lead mensuration costs an arm and a leg, and blood specimen often needs to be transferred to laboratory with good conditionsi to be analyzed, and at the problem that external paper disk method is deposited, the present invention proposes a new technical scheme that is more suitable for national conditions.
Summary of the invention
The objective of the invention is to: a kind of easy, quick, economic, children ' s lead poisoning screening method of tallying with the national condition is provided, makes the area that does not have respective detection equipment also can carry out the preventing and controlling of children ' s lead poisoning by the mode of mailing sample.
Purpose of the present invention can be achieved through the following technical solutions: a kind of method of paper disk method examination children ' s lead poisoning, step is divided into blood sample collection, the sample pre-service, the foundation of typical curve, test, be characterised in that: first step blood sample collection, successively children's is pointed and use the soap cotton balls, the 2%EDTA cotton balls, 75% cotton ball soaked in alcohol wiping, palm turned downwards to enjoin children's, all the other fingers of flexing, wait to point dry back with three shuttle pin acupunctures, extruding refers to abdomen gently, after forming enough big drop of blood, allow blood directly drip on filter paper, form diameter greater than the 0.6cm blood cake, whenever bleeding forms place's blood cake, drips two to three place's blood cakes on every filter paper, put into the polybag of handling through deionization after filter paper at room temperature dries, under 4-10 ℃ of environment temperature, preserve.It is convenient to have blood sampling, overlapping, blood coagulation that drop of blood is difficult for; The sample pre-service of second step, tying up to the blood cake center burrows and cuts out down diameter 0.6cm sequin and directly insert the nitric acid 200 μ l that add concentration 3-10% in the test tube, vibrate 5-10 second, make the scraps of paper fully be immersed in the salpeter solution, placed 16-24 hour under 4 ℃ of environment temperatures, vibrate once more after the taking-up 5-10 second, centrifugal treating is got supernatant and is tested then.It is few to have the number of oscillation, saves pretreatment time, and it is few to get specimen amount, is convenient to repeated test; The foundation of the 3rd step typical curve ties up in the scope of known blood lead concentration 0-500 μ g/L, chooses the 5-6 point, gets 30 μ l blood samples and drips on filter paper, and it is identical with test specimen that room temperature is dried the back, carries out testing after above-mentioned second step sample pre-service, makes curve.Make typical curve with whole blood, consistent with the blood specimen background of test, help to reduce error.Use Atomic Absorption Spectrometer (graphite furnace method) test at last.
The method of a kind of paper disk method examination children ' s lead poisoning of the present invention, described Atomic Absorption Spectrometer (graphite furnace method) test is on the technique scheme basis, Atomic Absorption Spectrometer wavelength 283.3nm, slit 0.7nm; The parameter of graphite furnace is dry 90-120 ℃, 30-40 second, ashing 600-700 ℃, 30 seconds; Atomization 1700-1800 ℃, 5 seconds, reading 2 seconds, each sample size of measuring is 20 μ l.It is few to have sample size, the characteristics that the test duration is short.
For validity and the feasibility of verifying paper disk method, can further obtain demonstration by following test result: one, the repeatability that the paper disk method blood lead is measured: get 30 parts of anticoagulations, drawing 30 μ l for every part drips on filter paper, repeat to do 4 parts of scraps of paper samples, measure once weekly, its result sees table 1 for details, the repeatability that the paper disk method blood lead is measured:
Table 1
Time example number blood lead (μ g/L) and venous blood related coefficient T value P value
First all 30 271.3+17.8 0.9391 14.4648＜0.01
Second all 30 285.6+18.9 0.9708 21.4106＜0.01
The 3rd all 30 280.7+20.4 0.9666 19.9635＜0.01
30 292.2+20.3 0.9553 17.0975＜0.01 around the
The blood lead concentration of venous blood is 270 μ g/L, and around first week to the, the related coefficient of blood lead is 0.9391-0.9708, and the good reproducibility that paper disk method test is fixed is described, and sample was measured in one month and do not influenced the result.Two, the heavy absorptivity of paper disk method: get 30 parts of anti-freezing venous blood of known lead concentration, draw 30 μ l for every part and drip on filter paper, measure the blood lead value of the scraps of paper, and contrast, and calculate heavy absorptivity with corresponding vein blood lead.The result is several 30 examples of example, heavy absorptivity scope 95.3-110.5%, average heavy absorptivity 98.8%.Three, the diffusibleness of different blood volumes on filter paper and the relation of blood lead content see table 2 for details, diffusibleness and the blood lead content of different blood volumes on filter paper:
Table 2
Blood volume example diameter area unit area contains the blood blood lead
μ l number (cm) (cm
2) amount (μ l/cm2) (μ g/L)
20 12 0.7805 0.4785 41.82+1.08* 96.09+21.23
30 12 0.9258 0.6732 44.59+1.43 111.09+36.59
50 12 1.1942 1.1201 44.70+1.94 112.08+52.75
The result shows, 20 μ l blood volumes are dripped on filter paper, its unit area contains the respective value that blood volume and blood lead concentration all are lower than 30 μ l and 50 μ l, and handling by statistics has marked difference (p＜0.01), and its unit area contains blood volume and the equal indifference of blood lead concentration (p＞0.05) between 30 μ l and the 50 μ l.Illustrate that (between the drop of blood 30 μ l-50 μ l, diffusibleness and the blood lead concentration of blood on filter paper is more constant within the specific limits.Four, the positive rate of paper disk method, false negative rate and false positive rate, in 158 routine children's, what the venous blood lead level was higher than 100 μ g/L, 200 μ g/L has 63 and 2 examples respectively, and scraps of paper rule has 55 and 2 examples respectively.Do positive rate, false negative rate and the false positive rate analysis of paper disk method examination children ' s lead poisoning by the standard of 100 μ g/L and 200 μ g/L, see Table 3, the positive rate of paper disk method, false negative rate and false positive rate:
Table 3
Positive rate % false negative rate % false positive rate % blood lead μ g/L
≥100 86.9 15.9 8.4
≥200 100 0 0
Though paper disk method exists certain false negative and false positive, in the error range (40 μ g/L) that the blood lead result generally acknowledges in the world.Five, the correlativity of venous blood and tip paper disk method blood lead concentration sees table 4 for details, the correlativity of venous blood and tip paper disk method blood lead concentration:
Table 4
Method example number blood lead μ g/L
Vein method 158 94.55+35.59
Paper disk method 158 92.75+33.87
Object is children's 158 examples in 2-7 year, gathers venous blood 1ml and puts into anticoagulant tube, immediately adopts finger tip blood on the scraps of paper.Venous blood adopts atomic absorption spectrography (AAS), and the processing of scraps of paper blood and detection method are the present invention, the correlation coefficient r of two kinds of its blood lead concentrations of assay method=0.8689, t=21.9636, p＜0.01.The further probatio inspectionem pecuoarem of above-mentioned test reliability of the present invention.Simultaneously, after the typical curve correction, the present invention also can be used for teenager, adult group.
Superiority of the present invention is: blood specimen is easily gathered, convenient transportation, the generaI investigation of blood lead also can be carried out in area to no checkout equipment, the result who detects meets internationally recognized critical field, establish the methodology basis for large-scale children ' s lead poisoning epidemiological study, the invention provides a kind of easy, quick, economic, children ' s lead poisoning screening method of tallying with the national condition.Below by embodiment in detail the present invention is described in detail.
Embodiment
Embodiment 1, the purpose of the method invention of paper disk method examination children ' s lead poisoning can be achieved through the following technical solutions: a kind of method of paper disk method examination blood lead, step is divided into blood sample collection, the sample pre-service, the foundation of typical curve, test, characteristics are: first step blood sample collection, successively children's is pointed and use the soap cotton balls, the 2%EDTA cotton balls, 75% cotton ball soaked in alcohol wiping, palm turned downwards to enjoin children's, all the other fingers of flexing, wait to point dry back with three shuttle pin acupunctures, extruding refers to abdomen gently, forms enough big drop of blood after, allow direct of blood on filter paper, form diameter greater than the 0.6cm blood cake, be advisable with 0.9-1.1cm, whenever bleeding forms place's blood cake, drips two to three place's blood cakes on every filter paper, put into the polybag of handling through deionization after filter paper at room temperature dries, in 4-10 ℃ of refrigerator, preserve.It is convenient to have blood sampling, overlapping, blood coagulation that drop of blood is difficult for; The sample pre-service of second step, tying up to the blood cake center burrows and cuts out down diameter 0.6cm sequin and directly insert in the round bottom plastic tube, the nitric acid 200 μ l that add concentration 5% then, oscillator vibration 10 seconds, the scraps of paper fully are immersed in the salpeter solution, in 4 ℃ of refrigerators, placed 16-24 hour, vibrated once more after the taking-up 10 seconds, 3000 rev/mins of centrifugal treating are 2 minutes then, and drawing supernatant is that available atomic absorption instrument (graphite furnace method) is measured.It is few to have the number of oscillation, saves pretreatment time, and it is few to get specimen amount, is convenient to the advantage of repeated test; The foundation of the 3rd step typical curve, the plumbous Quality Control sample that present embodiment selects for use the Center for Disease Control to provide in every month, in the blood lead concentration scope of 0-500 μ g/L, select 5 points, drip on filter paper with the microsyringe 30 μ l that suck blood, room temperature is dried, after burrowing the scraps of paper are placed the round bottom plastic tube, do identical sample pre-service simultaneously with test specimens.Test condition of the present invention is for adopting 4100 type Atomic Absorption Spectrometers of Perkin-Elmer company, wavelength 283.3nm, slit 0.70nm.The parameter of graphite furnace is dry 100 ℃, 30 seconds; 700 ℃ of ashing, 30 seconds; 1800 ℃ of atomization, 5 seconds, reading 2 seconds.Each sample size of measuring is 20 μ l.
Claims (2)
1, a kind of method of paper disk method examination children ' s lead poisoning, step be divided into blood sample collection, sample pre-service, typical curve foundation, test with Atomic Absorption Spectrometer, it is characterized in that: first step blood sample collection, get finger blood, enough big drop of blood is directly dripped on filter paper, form diameter greater than the 0.6cm blood cake, whenever bleed and form place's blood cake, drip two to three place's blood cakes on every filter paper, put into the polybag of handling through deionization after filter paper at room temperature dries, under 4-10 ℃ of environment temperature, preserve; The sample pre-service of second step, tying up to the blood cake center burrows and cuts out down diameter 0.6cm sequin and directly insert the nitric acid 200 μ l that add concentration 3-10% in the test tube, vibrate 5-10 second, make the scraps of paper fully be immersed in the salpeter solution, placed 16-24 hour under 4 ℃ of environment temperatures, vibrate once more after the taking-up 5-10 second, centrifugal treating is got supernatant and is tested then; The foundation of the 3rd step typical curve ties up in the blood lead concentration scope of known blood lead concentration 0-500 μ g/L selects the 5-6 point, gets 30 μ l blood samples and drips on filter paper, and it is identical with test specimen that room temperature is dried the back, carries out above-mentioned second and tests after going on foot the sample pre-service, makes curve.
2, the method for a kind of paper disk method examination children ' s lead poisoning according to claim 1 is characterized in that: Atomic Absorption Spectrometer wavelength 283.3nm, slit 0.7nm; The parameter of graphite furnace is dry 90-120 ℃, 30-40 second, ashing 600-700 ℃, 30 seconds; Atomization 1700-1800 ℃, 5 seconds, reading 2 seconds, each sample size 20 μ l that measure.
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|---|---|---|---|
| CNB001154117A CN1156707C (en) | 2000-04-19 | 2000-04-19 | Children's lead poisoning screening paper sheet method |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB001154117A CN1156707C (en) | 2000-04-19 | 2000-04-19 | Children's lead poisoning screening paper sheet method |
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| Publication Number | Publication Date |
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| CN1318744A CN1318744A (en) | 2001-10-24 |
| CN1156707C true CN1156707C (en) | 2004-07-07 |
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| CNB001154117A Expired - Fee Related CN1156707C (en) | 2000-04-19 | 2000-04-19 | Children's lead poisoning screening paper sheet method |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100460865C (en) * | 2006-02-22 | 2009-02-11 | 北京吉天仪器有限公司 | Method for measuring lead in blood by using hydride generating atomic spectrum method |
| AU2007327150B2 (en) * | 2006-12-01 | 2012-07-12 | Statens Serum Institut | Screening method using sample adsorption on filter paper |
| CN113686998A (en) * | 2021-09-28 | 2021-11-23 | 江苏晗睿达精准医学科技有限公司 | Method for determining trimethylamine oxide in dried blood spots by LC-MS/MS technology |
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