CN102636654A - Kit for determining concentration of human serum complement Clq and method thereof - Google Patents
Kit for determining concentration of human serum complement Clq and method thereof Download PDFInfo
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Abstract
The invention belongs to the field of biological engineering, provides a kit for determining the concentration of serum complement Clq, and solves the technical problem of complicated procedures, low accuracy and poor repeatability of immunodiffusion method and ELISA double-antibody sandwich technology adopted in the prior art to determine the concentration of complement Clq. The method involves the following two reagents: reagent I is composed of disodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-Na2 and TX-100, and reagent II is composed of disodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-Na2, TX-100, complement C1q antiserum or complement C1q monoclonal antibody. The invention also provides a method for determining the concentration of human serum complement Clq using the above kit. The kit and the method for determination of the concentration of complement C1q, provided by the invention, have the advantages of simple and convenient procedures, high accuracy and good repeatability, and are used for automated analyzers.
Description
Technical field
The invention belongs to bioengineering field, relate in particular to a kind of detection kit, particularly a kind of adopt immune turbidimetry and immune scattering turbidimetry detect in the human serum with arthral fluid in the kit and the method thereof of complement Clq concentration.
Background technology
Complement Clq is first composition of complement system Cl, is a giant molecule amount glycoprotein, and a Clq molecule is become by 18 polypeptied chains, and chemical composition is collagen protein molecular weight: 410KD.Available technology adopting SRID and ELISA double-antibody sandwich technical measurement complement Clq concentration, but its complicated steps, accuracy is not high.
Summary of the invention
The object of the present invention is to provide a kind of kit and method thereof that detects complement Clq concentration in the human serum; The kit of complement Clq concentration and method thereof will solve available technology adopting SRID and ELISA double-antibody sandwich technical measurement complement Clq concentration process complicacy, the technical matters that accuracy is not high in the described this detection human serum.
The invention provides a kind of kit that detects complement Clq concentration in the human serum, comprise two kinds of reagent, I reagent is by sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA
2Form with TX-100, in described I reagent, the mass concentration of described sodium hydrogen phosphate is 28.6g/L, and the mass concentration of described potassium dihydrogen phosphate is 2.7g/L, and the mass concentration of described PEG6000 is 50g/L, described EDTA-NA
2Mass concentration be 1g/L, the volumetric concentration of described TX-100 is 0.2ml/L, II reagent is by sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA
2, TX-100 and complement Clq antiserum or complement Clq monoclonal antibody form; In described II reagent; The mass concentration of described sodium hydrogen phosphate is 28.6g/L; The mass concentration of described potassium dihydrogen phosphate is 2.7g/L, and the mass concentration of described PEG6000 is 50g/L, described EDTA-NA
2Mass concentration be 1g/L, the volumetric concentration of described TX-100 is 0.2ml/L, the volumetric concentration of described complement Clq antiserum or complement Clq monoclonal antibody is 300ml/L.
Further, described complement Clq antiserum is goat-anti people complement Clq antiserum or mouse-anti people complement Clq antiserum, and described complement Clq monoclonal antibody is a mouse-anti people complement Clq monoclonal antibody.
Further, the purity of described TX-100 is 100%.
Further, sero-fast the tiring of the anti-people's complement of described rabbit Clq is 1: 64.
The present invention also provides a kind of method that detects complement Clq concentration in the human serum, adopts above-mentioned kit, comprises a step that adopts Biochemical Analyzer to measure in sample; Sample adopted in the step that Biochemical Analyzer measures at described one, sample is added I reagent R1 after, hatch 5min for 37 ℃; Read to survey the 1st sample A1, add II reagent R2, hatch 5min for 37 ℃; Read to survey the 2nd sample A2, sample A=sample A2-sample A1; Also comprise a step that adopts Biochemical Analyzer to measure calibration object, will calibrate at described one and adopt in the step that Biochemical Analyzer measures, will calibrate adding I reagent R1 after; Hatch 5min for 37 ℃; Read to survey the 1st calibration A1, add II reagent R2, hatch 5min for 37 ℃; Read to survey the 2nd calibration A2, calibration A=calibration A2-calibration A1;
The parameter of said determination is: 37 ℃ of temperature, predominant wavelength 340nm, 546nm, 600nm, commplementary wave length 700nm, sample or calibration object 3-4 μ l, R
1: 240 μ l, R
2: 60 μ l, the reaction time is 10 minutes.
Goat-anti people complement Clq antiserum of the present invention, mouse-anti people complement Clq antiserum, mouse-anti people complement Clq monoclonal antibody can be bought in market, also can be through conventional immunization method preparation.
Kit of the present invention and method are to adopt the measuring principle of immune turbidimetry and immune scattering turbidimetry; Promptly utilize the anti-people's complement of complement Clq antigen and rabbit Clq antiserum, goat-anti people complement Clq antiserum, mouse-anti people complement Clq antiserum, mouse-anti people complement Clq monoclonal antibody to combine; Form insolubilized immune complexes; It is muddy that reactant liquor is produced; Its turbidity height, the concentration that promptly penetrability reduces, absorbance increases complement Clq in the reflection human serum sample, the concentration of complement Clq can be calculated by the dose-effect curve that calibration object is done.
Adopting the concentration of the Clq that kit of the present invention and method record to can be used as scientific research uses with teaching.
The present invention compares with prior art, and its technical progress is significant.It is simple and convenient that kit of the present invention and method are used to measure the process of complement Clq concentration, and accuracy is high.
Description of drawings
Fig. 1 is the complement Clq concentration done by calibration object and the dose-effect curve figure of absorbance.
Embodiment
Embodiment 1 immune turbidimetry
When the IC in a branch of incident light irradiation liquid phase, a part of light is absorbed by the IC particle, a part of light transmission, and some light is by the IC KPT Scatter.Utilization is measured the ratio or the light absorption value of transmitted intensity and incident intensity than turbid appearance on the optical path direction of light source, judge the amount of determined antigen, and this method is called turbidimetry.
1.1 be suitable for instrument
Semi-automatic, automatic clinical chemistry analyzer.
1.2 analytical approach
The immunity turbidimetry.
1.3 performance requirement
1.3.1 reagent outward appearance
R1: achromaticity and clarification transparency liquid;
R2: faint yellow supernatant liquid.
1.3.2 reagent blank absorbance (A)
Absorbance (A): R
1+ R
2≤0.04A is (37 ℃ of temperature; Predominant wavelength 340nm (commplementary wave length 700nm)).
1.3.3 precision
1.3.3.1 withinrun precision
CV≤10％。
1.3.4 betweenrun precision
Relative extreme difference≤10%.
1.3.5 accuracy
Inaccuracy: in ± 10% the scope.
1.3.6 sensitivity for analysis
Absorbance (A)＞0.04A.
1.3.7 it is linear
In the 50mg/L-400mg/L scope, related coefficient (γ) >=0.9900.
1.3.8 stability
Reagent is stored to effective former and later two months in the end of term 2 ℃ of-8 ℃ of lucifuges, detects, and its quality meets the regulation of item.
2.1 testing conditions
2.1.1 Biochemical Analyzer (hereinafter to be referred as instrument)
Hitachi's 7060 automatic biochemistry analyzers.
2.1.2 operating ambient temperature
Room temperature 15-32 ℃.
Indoor humidity 45-85%RH.
2.1.3 main location parameter and operation steps
37 ℃ of temperature; Predominant wavelength 340nm (commplementary wave length 700nm), sample or calibration object 3-4 μ l; R
1: 240 μ l; R
2: 60 μ l, 10 minutes reaction time.
Method type: 2 end-point methods.
After sample or calibration object add reagent R1, hatch 5min for 37 ℃, read to survey the 1st point (A1), add reagent R2, hatch 5min for 37 ℃, read to survey the 2nd point (A2), sample A=A2-A1.
The result calculates:
2.1.4 calibration object
Used calibration object is the standard items that south, Yuhuan county, Zhejiang Province chemical reagent work produces in this standard.
2.2 reagent outward appearance
Visual observation agent box under the light, R1: achromaticity and clarification transparency liquid; R2: faint yellow supernatant liquid.
2.3 reagent blank absorbance
Get R
1240 μ l, adding distil water 4 μ l, put 37 ℃ hatch 5min after, add R
260 μ l, put 37 ℃ hatch 5min after, on 7060 automatic biochemistry analyzers, use the 340nm wavelength, measure the regulation that its absorbance should meet 1.3.2 item among the embodiment 1.
2.4 precision
2.4.1 withinrun precision test method
Under the instrument normal running conditions; Use with a collection of reagent; Follow-on test (about 200mg/L) sample 20 times; Calculate the mean value
and the standard deviation (SD) of its measured value; Calculate the value of the coefficient of variation (CV%) again by following formula, its result should meet the regulation of 1.3.3.1 item among the embodiment 1.
n
SD＝{∑(Xi-X)
2/(N-1)}
1/2
I＝1
In the formula: the SD-standard deviation;
The CV-coefficient of variation;
The average of
-n time measured value;
The measured value that Xi-is the i time;
N-measures number of times.
2.4.2 betweenrun precision test method
Under the instrument normal running conditions, get three lot number reagent, each lot number is got a cover.Measure (about 200mg/L) sample each 3 times respectively.Calculate every batch of average
of measuring the result and three crowdes of reagent mensuration results' grand mean
then and obtain the relative extreme difference (%) of the mensuration average of three lot number reagent according to formula, its result should meet the regulation of 1.3.4 item among the embodiment 1.
In the formula: maximal value during
is
, minimum value during
is
.
2.5 accuracy
Under the instrument normal running conditions, the standard items that reagent is produced with Zhejiang Yuhuan south chemical reagent work are measured the sample of concentration known after calibration, replication 10 times, and the relative deviation of the average of its measurement result should meet the regulation of 1.3.5 item among the embodiment 1.
RE%=absolute deviation/TV * 100%
Absolute deviation=testing result average-standard items target value
In the formula: the target value of TV-bioassay standard article
2.6 sensitivity for analysis
R
1Reagent 240 μ l add 50mg/L sample 4 μ l, put 37 ℃ hatch 5min after, add R
260 μ l, put 37 ℃ hatch 5min after, on 7060 automatic biochemistry analyzers,, survey absorbance with 340nm predominant wavelength (commplementary wave length 700nm), its result should meet the regulation of 1.3.6 item among the embodiment 1.
2.7 linear test
Sample or calibration object: select for use by 1 of embodiment 3 preparations
#, 2
#, 3
#, 4
#, and 5
#Each test concentrations sample or calibration object.
Determination step: under the instrument normal running conditions, after calibration, measure above five concentration samples or calibration object with reagent, each concentration samples or calibration object are measured respectively 3 times, and getting average is measured value Yi.
Calculate: calculate related coefficient γ by following formula, its result should meet the regulation of 1.3.7 item among the embodiment 1.
In the formula: γ-related coefficient
Xi-i
#The theoretical value of sample
Yi-i
#The measured value of sample
The numbering of i-variable concentrations test sample book
2.8 stability
Be taken at and be saved under the regulation storage requirement that the reagent in two months detects before and after effective end of term, its result should meet the regulation of 1.3.8 item among the embodiment 1.
Embodiment 2 immune scattering turbidimetrys
When the IC in a branch of incident light irradiation liquid phase, a part of light is absorbed by the IC particle, a part of light transmission, and some light is by the IC KPT Scatter.Measure scattered intensity at the certain angle (as 70 °) of light source light path, electric signal and scattered light intensity on the photoelectric cell are directly proportional, and convert the content of tested antigen to through micro computer, and this method is called the scattering turbidimetry method.
1.1 be suitable for instrument
Semi-automatic, automatic clinical chemistry analyzer.
1.2 analytical approach
Immunity scattering turbidimetry.
1.3 performance requirement
1.3.1 reagent outward appearance
R1: achromaticity and clarification transparency liquid;
R2: faint yellow supernatant liquid.
1.3.2 reagent blank absorbance (A)
Absorbance (A): R
1+ R
2≤0.04A is (37 ℃ of temperature; Predominant wavelength 340nm (commplementary wave length 700nm)).
1.3.3 precision
1.3.3.1 withinrun precision
CV≤10％。
1.3.4 betweenrun precision
Relative extreme difference≤10%.
1.3.5 accuracy
Inaccuracy: in ± 10% the scope.
1.3.6 sensitivity for analysis
Absorbance (A)＞0.04A.
1.3.7 it is linear
In the 50mg/L-400mg/L scope, related coefficient (γ) >=0.9900.
1.3.8 stability
Reagent is stored to effective former and later two months in the end of term 2 ℃ of-8 ℃ of lucifuges, detects, and its quality meets the regulation of item.
2.1 testing conditions
2.1.1 Biochemical Analyzer (hereinafter to be referred as instrument)
Hitachi's 7060 automatic biochemistry analyzers.
2.1.2 operating ambient temperature
Room temperature 15-32 ℃.
Indoor humidity 45-85%RH.
2.1.3 main location parameter and operation steps
37 ℃ of temperature; Predominant wavelength 340nm (commplementary wave length 700nm), sample or calibration object 3-4 μ l; R
1: 240 μ l; R
2: 60 μ l, 10 minutes reaction time.
Method type: 2 end-point methods.
After sample or calibration object add reagent R1, hatch 5min for 37 ℃, read to survey the 1st point (A1), add reagent R2, hatch 5min for 37 ℃, read to survey the 2nd point (A2), sample A=A2-A1.
The result calculates:
2.1.4 calibration object
Used calibration object is the standard items that south, Yuhuan county, Zhejiang Province chemical reagent work produces in this standard.
2.2 reagent outward appearance
Visual observation agent box under the light, R1: achromaticity and clarification transparency liquid; R2: faint yellow supernatant liquid.
2.3 reagent blank absorbance
Get R
1240 μ l, adding distil water 4 μ l, put 37 ℃ hatch 5min after, add R
260 μ l, put 37 ℃ hatch 5min after, on 7060 automatic biochemistry analyzers, use the 340nm wavelength, measure the regulation that its absorbance should meet 1.3.2 item among the embodiment 1.
2.4 precision
2.4.1 withinrun precision test method
Under the instrument normal running conditions; Use with a collection of reagent; Follow-on test (about 200mg/L) sample 20 times; Calculate the mean value
and the standard deviation (SD) of its measured value; Calculate the value of the coefficient of variation (CV%) again by following formula, its result should meet the regulation of 1.3.3.1 item among the embodiment 1.
n
SD＝{∑(Xi-X)
2/(N-1)}
1/2
I＝1
In the formula: the SD-standard deviation;
The CV-coefficient of variation;
The average of
-n time measured value;
The measured value that Xi-is the i time;
N-measurement number of times.
2.4.2 betweenrun precision test method
Under the instrument normal running conditions, get three lot number reagent, each lot number is got a cover.Measure (about 200mg/L) sample each 3 times respectively.Calculate every batch of average
of measuring the result and three crowdes of reagent mensuration results' grand mean
then and obtain the relative extreme difference (%) of the mensuration average of three lot number reagent according to formula, its result should meet the regulation of 1.3.4 item among the embodiment 1.
In the formula: maximal value during
is
, minimum value during
is
.
2.5 accuracy
Under the instrument normal running conditions, the standard items that reagent is produced with Zhejiang Yuhuan south chemical reagent work are measured the sample of concentration known after calibration, replication 10 times, and the relative deviation of the average of its measurement result should meet 1.3.5 among the embodiment 1
The regulation of item.
RE%=absolute deviation/TV * 100%
Absolute deviation=testing result average-standard items target value
In the formula: the target value of TV-bioassay standard article
2.6 sensitivity for analysis
R
1Reagent 240 μ l add 50mg/L sample 4 μ l, put 37 ℃ hatch 5min after, add R
260 μ l, put 37 ℃ hatch 5min after, on 7060 automatic biochemistry analyzers,, survey absorbance with 340nm predominant wavelength (commplementary wave length 700nm), its result should meet the regulation of 1.3.6 item among the embodiment 1.
2.7 linear test
Sample or calibration object: select for use by 1 of embodiment 3 preparations
#, 2
#, 3
#, 4
#, and 5
#Each test concentrations sample or calibration object.
Determination step: under the instrument normal running conditions, after calibration, measure above five concentration samples or calibration object with reagent, each concentration samples or calibration object are measured respectively 3 times, and getting average is measured value Yi.
Calculate: calculate related coefficient γ by following formula, its result should meet the regulation of 1.3.7 item among the embodiment 1.
In the formula: γ-related coefficient
Xi-i
#The theoretical value of sample
Yi-i
#The measured value of sample
The numbering of i-variable concentrations test sample book
2.8 stability
Be taken at and be saved under the regulation storage requirement that the reagent in two months detects before and after effective end of term, its result should meet the regulation of 1.3.8 item among the embodiment 1.
Embodiment 3
The preparation of reagent
Get the sample that a content is about 200mg/L, be sample 4
#, with sample 4
#The according to the form below method is mixed with 5 test sample books with physiological saline or pure water.(this sample is at present with join at present)
| 1 # | 2 # | 3 # | 4 # | 5 # | |
| Calibration object | - | 4μl | 2μl | 4μl | 8μl |
| Physiological saline | 4μl | 12μl | - | - | ?- |
| Dilution back sampling amount | - | 4μl | - | - | - |
| Concentration (mg/L) | 0 | 50 | 100 | 200 | 400 |
After sample or calibration object add reagent R1, hatch 5min for 37 ℃, read to survey the 1st point (A1), add reagent R2, hatch 5min for 37 ℃, read to survey the 2nd point (A2), sample A=A2-A1.
Instrument calculates as a result:
Through the mensuration of above-mentioned sample, obtained the dose-effect curve figure of complement Clq concentration and absorbance shown in Figure 1.
(Fig. 1 is the curve of being done according to the OD value of wavelength 340nm, the usefulness only for reference of wavelength 700nm.)
Pass through Fig. 1; Measured 500 routine normal persons' serum; Result such as following table are described; According to complement Clq normal reference value in serum of introducing in the Department of Medical Administration of the Ministry of Public Health " national clinical examination rules ": 157-237mg/L, the concentration that the Clq that adopts kit of the present invention and method mensuration is described is still accurately with reliably.
Complement Clq concentration (mg/L) in the 500 routine healthy subjects human serums
| Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | Numerical value |
| 1 | 179 | 9 | 196 | 17 | 196 | 25 | 221 | 33 | 206 | 41 | 195 |
| 2 | 174 | 10 | 199 | 18 | 178 | 26 | 205 | 34 | 195 | 42 | 205 |
| 3 | 166 | 11 | 201 | 19 | 221 | 27 | 192 | 35 | 206 | 43 | 200 |
| 4 | 198 | 12 | 193 | 20 | 207 | 28 | 196 | 36 | 182 | 44 | 198 |
| 5 | 206 | 13 | 194 | 21 | 197 | 29 | 213 | 37 | 198 | 45 | 215 |
| 6 | 195 | 14 | 203 | 22 | 194 | 30 | 206 | 38 | 196 | 46 | 161 |
| 7 | 216 | 15 | 221 | 23 | 184 | 31 | 191 | 39 | 194 | 47 | 180 |
| 8 | 194 | 16 | 196 | 24 | 207 | 32 | 192 | 40 | 226 | 48 | 196 |
| 49 | 205 | 78 | 195 | 107 | 196 | 136 | 206 | 165 | 196 | 194 | 193 |
| 50 | 200 | 79 | 194 | 108 | 194 | 137 | 200 | 166 | 192 | 195 | 203 |
| 51 | 160 | 80 | 206 | 109 | 196 | 138 | 192 | 167 | 194 | 196 | 191 |
| 52 | 194 | 81 | 187 | 110 | 199 | 139 | 196 | 168 | 195 | 197 | 200 |
| 53 | 206 | 82 | 198 | 111 | 199 | 140 | 197 | 169 | 217 | 198 | 198 |
| 54 | 184 | 83 | 225 | 112 | 200 | 141 | 195 | 170 | 198 | 199 | 183 |
| 55 | 163 | 84 | 217 | 113 | 196 | 142 | 196 | 171 | 196 | 200 | 226 |
| 56 | 206 | 85 | 193 | 114 | 195 | 143 | 198 | 172 | 193 | 201 | 195 |
| 57 | 207 | 86 | 195 | 115 | 196 | 144 | 199 | 173 | 188 | 202 | 198 |
| 58 | 205 | 87 | 204 | 116 | 194 | 145 | 232 | 174 | 198 | 203 | 194 |
| 59 | 198 | 88 | 196 | 117 | 196 | 146 | 193 | 175 | 195 | 204 | 206 |
| 60 | 195 | 89 | 198 | 118 | 201 | 147 | 194 | 176 | 194 | 205 | 198 |
| 61 | 230 | 90 | 196 | 119 | 195 | 148 | 196 | 177 | 229 | 206 | 198 |
| 62 | 197 | 91 | 195 | 120 | 191 | 149 | 163 | 178 | 195 | 207 | 191 |
| 63 | 194 | 92 | 182 | 121 | 193 | 150 | 195 | 179 | 204 | 208 | 195 |
| 64 | 195 | 93 | 207 | 122 | 195 | 151 | 196 | 180 | 199 | 209 | 193 |
| 65 | 195 | 94 | 198 | 123 | 179 | 152 | 172 | 181 | 198 | 210 | 195 |
| 66 | 198 | 95 | 217 | 124 | 197 | 153 | 192 | 182 | 183 | 211 | 161 |
| 67 | 205 | 96 | 203 | 125 | 197 | 154 | 203 | 183 | 195 | 212 | 199 |
| 68 | 204 | 97 | 192 | 126 | 166 | 155 | 229 | 184 | 196 | 213 | 219 |
| 69 | 187 | 98 | 196 | 127 | 195 | 156 | 197 | 185 | 198 | 214 | 195 |
| 70 | 228 | 99 | 229 | 128 | 194 | 157 | 226 | 186 | 205 | 215 | 192 |
| 71 | 199 | 100 | 197 | 129 | 196 | 158 | 198 | 187 | 206 | 216 | 187 |
| 72 | 196 | 101 | 192 | 130 | 196 | 159 | 196 | 188 | 196 | 217 | 196 |
| 73 | 198 | 102 | 195 | 131 | 195 | 160 | 200 | 189 | 201 | 218 | 193 |
| 74 | 196 | 103 | 181 | 132 | 193 | 161 | 192 | 190 | 233 | 219 | 206 |
| 75 | 184 | 104 | 186 | 133 | 201 | 162 | 184 | 191 | 226 | 220 | 197 |
| 76 | 202 | 105 | 194 | 134 | 193 | 163 | 188 | 192 | 188 | 221 | 194 |
| 77 | 196 | 106 | 198 | 135 | 228 | 164 | 204 | 193 | 194 | 222 | 195 |
| 223 | 207 | 252 | 204 | 281 | 193 | 310 | 182 | 339 | 165 | 368 | 198 |
| 224 | 228 | 253 | 216 | 282 | 207 | 311 | 196 | 340 | 196 | 369 | 219 |
| 225 | 199 | 254 | 196 | 283 | 205 | 312 | 189 | 341 | 202 | 370 | 192 |
| 226 | 200 | 255 | 207 | 284 | 191 | 313 | 206 | 342 | 216 | 371 | 221 |
| 227 | 196 | 256 | 199 | 285 | 196 | 314 | 198 | 343 | 184 | 372 | 196 |
| 228 | 198 | 257 | 192 | 286 | 221 | 315 | 200 | 344 | 195 | 373 | 206 |
| 229 | 197 | 258 | 222 | 287 | 195 | 316 | 229 | 345 | 194 | 374 | 196 |
| 230 | 196 | 259 | 201 | 288 | 173 | 317 | 194 | 346 | 183 | 375 | 192 |
| 231 | 197 | 260 | 194 | 289 | 198 | 318 | 186 | 347 | 196 | 376 | 189 |
| 232 | 196 | 261 | 206 | 290 | 191 | 319 | 191 | 348 | 203 | 377 | 203 |
| 233 | 195 | 262 | 171 | 291 | 184 | 320 | 163 | 349 | 196 | 378 | 196 |
| 234 | 197 | 263 | 192 | 292 | 217 | 321 | 197 | 350 | 201 | 379 | 197 |
| 235 | 219 | 264 | 199 | 293 | 182 | 322 | 231 | 351 | 192 | 380 | 196 |
| 236 | 194 | 265 | 206 | 294 | 183 | 323 | 190 | 352 | 201 | 381 | 190 |
| 237 | 199 | 266 | 194 | 295 | 186 | 324 | 189 | 353 | 199 | 382 | 179 |
| 238 | 193 | 267 | 207 | 296 | 200 | 325 | 187 | 354 | 198 | 383 | 192 |
| 239 | 196 | 268 | 196 | 297 | 199 | 326 | 192 | 355 | 194 | 384 | 190 |
| 240 | 201 | 269 | 198 | 298 | 182 | 327 | 196 | 356 | 197 | 385 | 196 |
| 241 | 198 | 270 | 193 | 299 | 233 | 328 | 196 | 357 | 195 | 386 | 195 |
| 242 | 200 | 271 | 171 | 300 | 189 | 329 | 191 | 358 | 162 | 387 | 183 |
| 243 | 196 | 272 | 203 | 301 | 159 | 330 | 160 | 359 | 209 | 388 | 208 |
| 244 | 163 | 273 | 189 | 302 | 194 | 331 | 196 | 360 | 219 | 389 | 198 |
| 245 | 196 | 274 | 195 | 303 | 201 | 332 | 188 | 361 | 233 | 390 | 194 |
| 246 | 227 | 275 | 206 | 304 | 182 | 333 | 194 | 362 | 221 | 391 | 203 |
| 247 | 159 | 276 | 219 | 305 | 166 | 334 | 198 | 363 | 190 | 392 | 220 |
| 248 | 191 | 277 | 207 | 306 | 184 | 335 | 201 | 364 | 213 | 393 | 195 |
| 249 | 195 | 278 | 196 | 307 | 196 | 336 | 196 | 365 | 196 | 394 | 180 |
| 250 | 209 | 279 | 206 | 308 | 201 | 337 | 195 | 366 | 184 | 395 | 159 |
| 251 | 226 | 280 | 194 | 309 | 184 | 338 | 205 | 367 | 229 | 396 | 204 |
| 397 | 192 | 421 | 199 | 445 | 197 | 469 | 199 | 493 | 203 | 397 | 192 |
| 398 | 195 | 422 | 197 | 446 | 196 | 470 | 184 | 494 | 233 | 398 | 195 |
| 399 | 197 | 423 | 182 | 447 | 196 | 471 | 191 | 495 | 172 | 399 | 197 |
| 400 | 195 | 424 | 205 | 448 | 201 | 472 | 197 | 496 | 224 | 400 | 195 |
| 401 | 193 | 425 | 197 | 449 | 178 | 473 | 196 | 497 | 160 | ||
| 402 | 187 | 426 | 203 | 450 | 207 | 474 | 204 | 498 | 164 | ||
| 403 | 191 | 427 | 190 | 451 | 213 | 475 | 205 | 499 | 184 | ||
| 404 | 181 | 428 | 195 | 452 | 164 | 476 | 196 | 500 | 159 | ||
| 405 | 197 | 429 | 161 | 453 | 229 | 477 | 180 | ||||
| 406 | 232 | 430 | 181 | 454 | 192 | 478 | 194 | ||||
| 407 | 210 | 431 | 198 | 455 | 172 | 479 | 196 | ||||
| 408 | 182 | 432 | 191 | 456 | 200 | 480 | 193 | ||||
| 409 | 179 | 433 | 170 | 457 | 194 | 481 | 196 |
| 410 | 188 | 434 | 233 | 458 | 181 | 482 | 177 | ||||
| 411 | 209 | 435 | 193 | 459 | 178 | 483 | 196 | ||||
| 412 | 196 | 436 | 232 | 460 | 171 | 484 | 229 | ||||
| 413 | 180 | 437 | 204 | 461 | 189 | 485 | 198 | ||||
| 414 | 173 | 438 | 230 | 462 | 202 | 486 | 196 | ||||
| 415 | 194 | 439 | 159 | 463 | 166 | 487 | 163 | ||||
| 416 | 164 | 440 | 174 | 464 | 208 | 488 | 160 | ||||
| 417 | 200 | 441 | 184 | 465 | 211 | 489 | 233 | ||||
| 418 | 189 | 442 | 164 | 466 | 195 | 490 | 160 | ||||
| 419 | 173 | 443 | 189 | 467 | 196 | 491 | 191 | ||||
| 420 | 181 | 444 | 193 | 468 | 199 | 492 | 161 |
Claims (5)
1. a kit that detects complement Clq concentration in the human serum comprises two kinds of reagent, and it is characterized in that: I reagent is by sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA
2Form with TX-100, in described I reagent, the mass concentration of described sodium hydrogen phosphate is 28.6g/L, and the mass concentration of described potassium dihydrogen phosphate is 2.7g/L, and the mass concentration of described PEG6000 is 50g/L, described EDTA-NA
2Mass concentration be 1g/L, the volumetric concentration of described TX-100 is 0.2ml/L, II reagent is by sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA
2, TX-100 and C1Q antiserum or C1Q monoclonal antibody form; In described II reagent; The mass concentration of described sodium hydrogen phosphate is 28.6g/L; The mass concentration of described potassium dihydrogen phosphate is 2.7g/L, and the mass concentration of described PEG6000 is 50g/L, described EDTA-NA
2Mass concentration be 1g/L, the volumetric concentration of described TX-100 is 0.2ml/L, the volumetric concentration of described C1Q antiserum or C1Q monoclonal antibody is 300ml/L.
2. a kind of kit that detects complement Clq concentration in the human serum as claimed in claim 1; It is characterized in that: described C1Q antiserum is goat-anti people C1Q antiserum or mouse-anti people C1Q antiserum, and described C1Q monoclonal antibody is a mouse-anti people C1Q monoclonal antibody.
3. a kind of kit that detects complement Clq concentration in the human serum as claimed in claim 1, it is characterized in that: the purity of described TX-100 is 100%.
4. a kind of kit that detects complement Clq concentration in the human serum as claimed in claim 1 is characterized in that: sero-fast the tiring of the anti-people's C1Q of described rabbit is 1:64.
5. a method that detects complement Clq concentration in the human serum is characterized in that adopting the described kit of claim 1, comprises a step that adopts Biochemical Analyzer to measure in sample; Sample adopted in the step that Biochemical Analyzer measures at described one, sample is added I reagent R1 after, hatch 5min for 37 ℃; Read to survey the 1st sample A1, add II reagent R2, hatch 5min for 37 ℃; Read to survey the 2nd sample A2, sample A=sample A2-sample A1; Also comprise a step that adopts Biochemical Analyzer to measure calibration object, will calibrate at described one and adopt in the step that Biochemical Analyzer measures, will calibrate adding I reagent R1 after; Hatch 5min for 37 ℃; Read to survey the 1st calibration A1, add II reagent R2, hatch 5min for 37 ℃; Read to survey the 2nd calibration A2, calibration A=calibration A2-calibration A1;
Sample A2-sample A1
Complement Clq concentration (mg/L)=
* calibration object concentration (mg/L)
Calibration A2-calibration A1
The parameter of said determination is: 37 ℃ of temperature, predominant wavelength 340nm, 546nm, 600nm, commplementary wave length 700nm, sample or calibration object 3-4 μ l, R
1: 240 μ l, R
2: 60 μ l, the reaction time is 10 minutes.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012101336190A CN102636654A (en) | 2012-04-28 | 2012-04-28 | Kit for determining concentration of human serum complement Clq and method thereof |
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| CN2012101336190A CN102636654A (en) | 2012-04-28 | 2012-04-28 | Kit for determining concentration of human serum complement Clq and method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103645331A (en) * | 2013-12-24 | 2014-03-19 | 上海北加生化试剂有限公司 | Kit and method for detecting fibronectin concentration in human urine |
| CN107207587A (en) * | 2014-11-05 | 2017-09-26 | 安尼艾克松股份有限公司 | Humanization anticomplement factor C1Q antibody and its application |
| CN108169145A (en) * | 2017-11-17 | 2018-06-15 | 安徽伊普诺康生物技术股份有限公司 | A kind of kit for measuring serum complement C1q and its preparation application method |
| CN109752332A (en) * | 2017-11-07 | 2019-05-14 | 重庆中元汇吉生物技术有限公司 | A kind of C1Q detection kit |
| CN111141914A (en) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | N-telopeptide precursor detection kit and preparation method thereof |
| CN114137220A (en) * | 2021-10-22 | 2022-03-04 | 苏州普瑞斯生物科技有限公司 | Production process of complement C1q detection reagent by immunoturbidimetry |
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| WO2001073437A2 (en) * | 2000-03-27 | 2001-10-04 | Diagen Corporation | Antigen-specific enzyme-linked immunosorbent assay |
| US20040248156A1 (en) * | 2001-12-03 | 2004-12-09 | Tianhua Hu | Methods and materials relating to novel C1q domain-containing polypeptides and polynucleotides |
| CN102323427A (en) * | 2011-08-09 | 2012-01-18 | 上海北加生化试剂有限公司 | Kit and method for detecting concentration of complement Clq in human serum |
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2012
- 2012-04-28 CN CN2012101336190A patent/CN102636654A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001073437A2 (en) * | 2000-03-27 | 2001-10-04 | Diagen Corporation | Antigen-specific enzyme-linked immunosorbent assay |
| US20040248156A1 (en) * | 2001-12-03 | 2004-12-09 | Tianhua Hu | Methods and materials relating to novel C1q domain-containing polypeptides and polynucleotides |
| CN102323427A (en) * | 2011-08-09 | 2012-01-18 | 上海北加生化试剂有限公司 | Kit and method for detecting concentration of complement Clq in human serum |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103645331A (en) * | 2013-12-24 | 2014-03-19 | 上海北加生化试剂有限公司 | Kit and method for detecting fibronectin concentration in human urine |
| CN107207587A (en) * | 2014-11-05 | 2017-09-26 | 安尼艾克松股份有限公司 | Humanization anticomplement factor C1Q antibody and its application |
| CN107207587B (en) * | 2014-11-05 | 2022-04-19 | 安尼艾克松股份有限公司 | Humanized anti-complement factor C1Q antibodies and uses thereof |
| CN109752332A (en) * | 2017-11-07 | 2019-05-14 | 重庆中元汇吉生物技术有限公司 | A kind of C1Q detection kit |
| CN108169145A (en) * | 2017-11-17 | 2018-06-15 | 安徽伊普诺康生物技术股份有限公司 | A kind of kit for measuring serum complement C1q and its preparation application method |
| CN111141914A (en) * | 2020-01-02 | 2020-05-12 | 四川纳海川生物科技有限公司 | N-telopeptide precursor detection kit and preparation method thereof |
| CN114137220A (en) * | 2021-10-22 | 2022-03-04 | 苏州普瑞斯生物科技有限公司 | Production process of complement C1q detection reagent by immunoturbidimetry |
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