CN102650641B - Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry - Google Patents
Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry Download PDFInfo
- Publication number
- CN102650641B CN102650641B CN201210133492.2A CN201210133492A CN102650641B CN 102650641 B CN102650641 B CN 102650641B CN 201210133492 A CN201210133492 A CN 201210133492A CN 102650641 B CN102650641 B CN 102650641B
- Authority
- CN
- China
- Prior art keywords
- sample
- reagent
- concentration
- calibration
- complement
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the field of biological engineering, and provides a kit for detecting C1q concentration of a complement in blood serum with a nanosized-latex enhancing immune turbidimetry, solving the problems of complex steps, low accuracy and poor repeatability in the prior art adopting an immunodiffusion method and an ELISA (Enzyme-Linked Immuno Sorbent Assay) double-antibody sandwich technology to detect the C1q concentration of the complement. The kit comprises two reagents, wherein a reagent I is prepared from disodium hydrogen phosphate, monopotassium phosphate, PEG6000, EDTA-NA2 and TX-100; and a reagent II is prepared from suspensions of rabbit anti-human complement C1q antiserum or goat anti-human complement C1q antiserum or rat anti-human complement C1q antiserum or rat anti-human complement C1q monoclonal antibodies crosslinking on latex particles. The invention also provides a method using the kit to detect the C1q concentration of the complement in human blood serum. The kit and the method have simpleness and convenience in steps for detecting the C1q concentration of the complement, high degree of accuracy and good repeatability and are used for automatic analyzers.
Description
Technical field
The invention belongs to bioengineering field, relate in particular to a kind of detection kit, particularly a kind of kit and method thereof of strengthening complement Clq concentration in nano rubber latex immunoturbidimetry detection human serum and in arthral fluid of adopting.
Background technology
Complement Clq is the 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 technology are measured complement Clq concentration, but its step complexity, and accuracy is not high.
Summary of the invention
The object of the present invention is to provide a kind of kit and method that strengthens nano rubber latex immunoturbidimetry detection complement Clq concentration, described this enhancing nano rubber latex immunoturbidimetry detects the kit of complement Clq concentration and method will solve available technology adopting SRID and ELISA double-antibody sandwich technology is measured complement Clq concentration process complexity, the technical matters that accuracy is not high.
The invention provides a kind of kit that strengthens nano rubber latex immunoturbidimetry detection complement Clq concentration, comprise two kinds of reagent, it is characterized in that: I reagent is by sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000-8000, EDTA-NA2 and TX-100 composition, in I 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, the mass concentration of described PEG6000 is 50g/L, the mass concentration of described EDTA-NA2 is 1g/L, the volumetric concentration of described TX-100 is 0.2ml/L, II reagent is cross-linked suspension on latex particle by rabbit anti-human complement Clq antiserum or goat-anti people complement Clq antiserum or mouse-anti people complement Clq antiserum or mouse-anti people complement Clq monoclonal antibody and forms.
Further, described II reagent is to react rabbit anticomplement Clq antibody or goat-anti complement Clq antibody or mouse-anti complement Clq antibody by carbodiimide, or mouse-anti people complement Clq monoclonal antibody, IgG are covalently bound on latex particle.
Further, the purity of described TX-100 is 100%.
Further, sero-fast the tiring of the anti-human complement Clq of described rabbit is 1: 64.
The present invention also provides a kind of method that detects concentration of complement Clq in human serum, adopt above-mentioned kit, comprise a step that adopts Biochemical Analyzer to measure in sample, sample is adopted in the step that Biochemical Analyzer measures at described one, sample is added after I reagent R1, hatch 5min for 37 DEG C, read to survey the 1st sample A1, add II reagent R2, hatch 5min for 37 DEG C, 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, in a described step that calibration employing Biochemical Analyzer is measured, calibration is added after I reagent R1, hatch 5min for 37 DEG C, read to survey the 1st calibration A1, add II reagent R2, hatch 5min for 37 DEG C, read to survey the 2nd calibration A2, calibration A=calibration A2-calibration A1;
Result is calculated:
The parameter of said determination is: 37 DEG C of temperature, predominant wavelength 546nm or 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.
Rabbit of the present invention anti-human complement Clq antiserum or goat-anti people complement Clq antiserum or mouse-anti people complement Clq antiserum or mouse-anti people complement Clq monoclonal antibody can be bought in market, also can be by conventional immunization method preparation.
Kit of the present invention and method are to adopt the measuring principle that strengthens nano rubber latex immunoturbidimetry, the anti-human complement Clq of rabbit antiserum, goat-anti people complement Clq antiserum, mouse-anti people complement Clq antiserum, the mouse-anti people complement Clq monoclonal antibody of utilizing complement Clq antigen and be cross-linked on latex particle combine, form insolubilized immune complexes, reactant liquor is produced muddy, its turbidity height, be the concentration that penetrability reduces, absorbance increases complement Clq in reflection human serum sample, the dose-effect curve that the concentration of complement Clq can be done by calibration object calculates.
Adopt the concentration of the CLq that kit of the present invention and method record to can be used as scientific research and teaching is used.
The present invention compares with prior art, and its technical progress is significant.Kit of the present invention and method are simple and convenient for measuring the process of complement Clq concentration, and accuracy is high.
Brief description of the drawings
Fig. 1 is the complement Clq concentration done by calibration object and the dose-effect curve figure of absorbance.
Embodiment
1.1 are suitable for instrument
Semi-automatic, automatic clinical chemistry analyzer.
1.2 analytical approach
Strengthen nanometer latex immunoturbidimetry.
1.3 performance requirement
1.3.1 reagent outward appearance
R1: achromaticity and clarification transparency liquid;
R2: faint yellow supernatant liquid or latex suspension.
1.3.2 reagent blank absorbance (A)
Absorbance (A): R
1+ R
2≤ 0.04A is (37 DEG C of temperature; Predominant wavelength 546nm, 600nm (commplementary wave length 700nm)).
1.3.3 precision
1.3.3.1 withinrun precision
CV≤10%。
1.3.4 betweenrun precision
Extreme difference≤10% relatively.
1.3.5 accuracy
Inaccuracy: in ± 10% scope.
1.3.6 sensitivity for analysis
Absorbance (A) > 0.04A.
1.3.7 linear
Within the scope of 50mg/L-400mg/L, related coefficient (γ) >=0.9900.
1.3.8 stability
Reagent, detects to effective former and later two months in the end of term 2 DEG C of-8 DEG C of lucifuges storages, and its quality meets the regulation of item.
Experimental technique
Embodiment 2
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 DEG C.
Indoor humidity 45-85%RH.
2.1.3 main location parameter and operation steps
37 DEG C of temperature; Predominant wavelength 546nm, 600nm (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 reagent adding R1, hatch 5min for 37 DEG C, read to survey the 1st point (A1), add reagent R2, hatch 5min for 37 DEG C, read to survey the 2nd point (A2), sample A=A2-A1.
Result is calculated:
2.1.4 calibration object
In this standard, calibration object used is the standard items that south, Yuhuan county of Zhejiang Province chemical reagent work produces.
2.2 reagent outward appearances
Visual observation agent box under light, R1: achromaticity and clarification transparency liquid; R2: faint yellow supernatant liquid.
2.3 reagent blank absorbances
Get R
1240 μ l, adding distil water 4 μ l, put 37 DEG C and hatch after 5min, add R
260 μ l, put 37 DEG C and hatch after 5min, on 7060 automatic biochemistry analyzers, with 546nm, 600nm wavelength, measure the regulation that its absorbance should meet 1.3.2 item in embodiment 1.
2.4 precision
2.4.1 withinrun precision test method
Under instrument normal running conditions, to use with a collection of reagent, follow-on test (about 200mg/L) sample 20 times, calculates the mean value of its measured value
and standard deviation (SD), then calculate as follows the value of the coefficient of variation (CV%), its result should meet the regulation of 1.3.3.1 item in embodiment 1.
n
SD={∑(Xi-X)
2/(N-1)}
1/2
I=1
In formula: SD-standard deviation;
The CV-coefficient of variation;
The measured value that Xi-is the i time;
N-measures number of times.
2.4.2 betweenrun precision test method
Under instrument normal running conditions, get three lot number reagent, each lot number is got a set of.Measure respectively each 3 times of (about 200mg/L) sample.Then calculate the average of every batch of measurement result
grand mean with three batches of reagent measurement results
the relative extreme difference (%) of obtaining according to the following formula the mensuration average of three lot number reagent, its result should meet the regulation of 1.3.4 item in embodiment 1.
2.5 accuracy
Under instrument normal running conditions, the standard items that reagent is produced with Zhejiang Yuhuan south chemical reagent work, after calibration, are measured the sample of concentration known, replication 10 times, and the relative deviation of the average of its measurement result should meet the regulation of 1.3.5 item in embodiment 1.
RE%=absolute deviation/TV × 100%
Absolute deviation=testing result average-standard items target value
In formula: the target value of TV-bioassay standard product
2.6 sensitivity for analysis
R
1reagent 240 μ l, add 50mg/L sample 4 μ l, put 37 DEG C and hatch after 5min, add R
260 μ l, put 37 DEG C and hatch after 5min, and on 7060 automatic biochemistry analyzers, with 546nm, 600nm, predominant wavelength (commplementary wave length 700nm), surveys absorbance, and its result should meet the regulation of 1.3.6 item in embodiment 1.
2.7 linear test
Sample or calibration object: select prepare by embodiment 31
#, 2
#, 3
#, 4
#, and 5
#each test concentrations sample or calibration object.
Determination step: under 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 as follows related coefficient γ, its result should meet the regulation of 1.3.7 item in embodiment 1.
In 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
5.8 stability
Be taken at and under regulation storage requirement, be saved to the reagent in two months before and after effective end of term and detect, its result should meet the regulation of 1.3.8 item in embodiment 1.
Embodiment 3
The preparation of reagent
Getting the sample that a content is about 200mg/L, is sample 4
#, by sample 4
#according to the form below method is mixed with 5 test sample books with physiological saline or pure water.(this sample matching while using)
1 # | 2 # | 3 # | 4 # | 5 # | |
Calibration object | - | 4μl | 2μl | 4μl | 8μl |
Physiological saline | 4μl | 12μl | - | - | - |
Sampling amount after dilution | - | 4μl | - | - | - |
Concentration (mg/L) | 0 | 50 | 100 | 200 | 400 |
After sample or calibration object reagent adding R1, hatch 5min for 37 DEG C, read to survey the 1st point (A1), add reagent R2, hatch 5min for 37 DEG C, read to survey the 2nd point (A2), sample A=A2-A1.
Result instrument calculates:
By the mensuration of above-mentioned sample, obtain the dose-effect curve figure of the complement Clq concentration shown in Fig. 1 and absorbance.
(Fig. 1 is the curve doing according to the OD value of wavelength 600nm, the use only for reference of commplementary wave length 700nm.)
Pass through Fig. 1, measure 500 routine normal persons' serum, result is described as following table, complement Clq normal reference value in serum according to introducing in Department of Medical Administration of the Ministry of Public Health " national clinical examination code ": 157-237mg/L, the concentration that the Clq that adopts kit of the present invention and method mensuration be described still accurately and reliably.
500 routine Healthy People concentration of complement Clq in human serums (mg/L)
Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | Numerical value | Number | |
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 (2)
1. one kind strengthens the kit of nano rubber latex immunoturbidimetry detection complement Clq concentration, comprise two kinds of reagent, it is characterized in that: I reagent is made up of sodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000-8000, EDTA-NA2 and TX-100, in I 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, II reagent is cross-linked suspension on latex particle by the anti-human C1Q antiserum of rabbit or goat-anti people C1Q antiserum or mouse-anti people C1Q antiserum or mouse-anti people C1Q monoclonal antibody and forms, described II reagent is to react the antibody by rabbit anticomplement C1q by carbodiimide, or goat-anti C1Q antibody, or mouse-anti C1Q antibody, or mouse-anti people C1Q monoclonal antibody, IgG is covalently bound on latex particle, the purity of described TX-100 is 100%, the anti-human C1Q of described rabbit is sero-fast tires as 1:64.
2. one kind is detected the method for concentration of complement Clq in human serum, it is characterized in that adopting kit claimed in claim 1, comprise a step that adopts Biochemical Analyzer to measure in sample, sample is adopted in the step that Biochemical Analyzer measures at described one, sample is added after I reagent R1, hatch 5min for 37 DEG C, read to survey the 1st sample A1, add II reagent R2, hatch 5min for 37 DEG C, 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, in a described step that calibration employing Biochemical Analyzer is measured, calibration is added after I reagent R1, hatch 5min for 37 DEG C, read to survey the 1st calibration A1, add II reagent R2, hatch 5min for 37 DEG C, 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 DEG C of temperature, predominant wavelength 546nm or 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.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210133492.2A CN102650641B9 (en) | 2012-04-28 | Strengthen kit and the method that nano rubber latex immunoturbidimetry detects complement Clq concentration |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210133492.2A CN102650641B9 (en) | 2012-04-28 | Strengthen kit and the method that nano rubber latex immunoturbidimetry detects complement Clq concentration |
Publications (3)
Publication Number | Publication Date |
---|---|
CN102650641A CN102650641A (en) | 2012-08-29 |
CN102650641B true CN102650641B (en) | 2014-06-18 |
CN102650641B9 CN102650641B9 (en) | 2016-05-18 |
Family
ID=
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007043582A1 (en) * | 2005-10-11 | 2007-04-19 | Sysmex Corporation | Method for determination of sars virus nucleocapsid protein, reagent kit for the determination, test device, monoclonal antibody directed against sars virus nucleocapsid protein, and hybridoma capable of producing the monoclonal antibody |
CN101968492A (en) * | 2010-10-29 | 2011-02-09 | 厦门大学附属中山医院 | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof |
CN102253217A (en) * | 2011-04-07 | 2011-11-23 | 武汉生之源生物科技有限公司 | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein |
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 |
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007043582A1 (en) * | 2005-10-11 | 2007-04-19 | Sysmex Corporation | Method for determination of sars virus nucleocapsid protein, reagent kit for the determination, test device, monoclonal antibody directed against sars virus nucleocapsid protein, and hybridoma capable of producing the monoclonal antibody |
CN101968492A (en) * | 2010-10-29 | 2011-02-09 | 厦门大学附属中山医院 | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof |
CN102253217A (en) * | 2011-04-07 | 2011-11-23 | 武汉生之源生物科技有限公司 | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein |
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 |
Also Published As
Publication number | Publication date |
---|---|
CN102650641A (en) | 2012-08-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102628867B (en) | Double antibody latex intensified Retinal-binding protein detection kit | |
CN104395728B (en) | Improve sensitivity and the dynamic range of photometry by generating multiple calibration curves | |
CN102253217B (en) | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein | |
CN103604930B (en) | A kind of lipoprotein (a) detection kit | |
CN105510604A (en) | Method for improving sensitivity and linearity of latex reagent | |
CN103852584B (en) | A kind of latex enhancing immune of quantitative detection C peptide is than turbid kit | |
CN107402308A (en) | Immue quantitative detection reagent boxes of IL 6 and preparation method thereof | |
CN105352958A (en) | Detection reagent kit for overall 25-hydroxy-vitamin-D | |
CN102590497A (en) | Cysteine protease inhibitor C test kit | |
CN105158476A (en) | Full-scale range C-reactive protein latex-enhanced immunoturbidimetry detection kit | |
CN103364568A (en) | Laminin nano-magnetic particle chemiluminescent immunity quantitative detection kit and preparation method thereof | |
CN101833009A (en) | Double antibody complex retinol-binding protein assay kit | |
CN104711279B (en) | It is prepared by a kind of VEGF detection kit and its raw material | |
CN108613977B (en) | N-terminal brain natriuretic peptide precursor detection kit | |
Bocková et al. | Surface plasmon resonance biosensor for detection of pregnancy associated plasma protein A2 in clinical samples | |
CN102636654A (en) | Kit for determining concentration of human serum complement Clq and method thereof | |
CN104849473A (en) | Microalbuminuria detection kit and preparation thereof | |
CN103033619A (en) | Protein chip reagent kit and method for comprehensively detecting lung cancer marker | |
CN107957495A (en) | A kind of CK-MB detection kits and its application method | |
CN106324254A (en) | Anti-insulin antibody detection kit and detection method thereof | |
CN111781372A (en) | Alpha 1-AT immunoturbidimetry detection kit based on mixed antibody and preparation and use methods thereof | |
CN103675299A (en) | Kit and method for detecting concentration of fibronectin in urine | |
CN102866256A (en) | Detection method and detection reagent for hypersensitive C reactive protein | |
CN110045129B (en) | Method for detecting human soluble asialoglycoprotein receptor | |
CN107942068A (en) | β2Microglobulin assay kit |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C53 | Correction of patent for invention or patent application | ||
CI01 | Correction of invention patent gazette |
Correction item: Denomination of Invention Correct: Correct False: Error Number: 25 Volume: 30 |
|
CI03 | Correction of invention patent |
Correction item: Denomination of Invention|Claims|Description Correct: Correct False: Error Number: 25 Page: full text Volume: 30 |
|
ERR | Gazette correction |