APPLICATION OF THE PARALLEL LINE BIOASSAY METHOD TO QUANTITATIVE DETERMINATION OF HBs ANTIGEN IN RADIOIMMUNOASSAY
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Abstract:
Precise and reproducible quantitative determination of hepatitis B surface antigen by applying the parallel line bioassay method was proposed. In this method, the value of any given sample can be expressed in terms of a relative value to a fixed standard or reference preparation.Decrease in antigenic reactivity due to the lowered protein content could be prevented by using 0.02% human albumin in physiological saline solution as the diluent. The use of such stabilizer (s) should be considered in the dilution or storage of samples containing HBs antigen.Keywords:
Diluent
Dilution
Serial dilution
Sephadex
Colony-stimulating factor
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Intradermal injection
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Objective To discuss the HBsAg applicable diluent and optimization dilution multiple with i2000 Chemoluminescence Analyzer and provide evidence for diagnosis and therapy of HBV infection.Methods(1) The choice of dilution multiple: The optimization dilution multiple was determined by analyzing statistically frequency of different dilution multiple from 1 192 results which were detected with dilution method suggested by manufacturer in our Clinical Laboratory.(2) The choice of applicable diluent: 30 distinct patient serum(HBsAg250 IU/mL)were detected after 50,100,500 fold dilution with manufacturer diluent,mixture negative serum,normal sodium and 10% calf serum.The suitable diluent and dilution multiple were determined by analyzing statistically test results of manufacturer diluent,mixture negative serum,normal sodium and 10% calf serum.Results(1) In 1 192 diluted samples,95% of the test results were within 50 fold dilution range;98% of the test results were in 100 fold dilution range;99% of the test results were in the range of 500 fold dilution range.(2)There were no significantly different between the detection results of manufacturer diluent and those of mixture negative serum,normal sodium and 10% calf serum.(P0.05)Conclusion When HBsAg250 IU/mL,100 fold dilution is selected firstly.As HBsAg250 IU/mL after 100 fold dilution,500 fold dilution is selected and detected.The manufacturer diluent,mixture negative serum,normal sodium and 10% calf serum are as applicable diluent.
Diluent
Dilution
Serial dilution
Isotope dilution
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Dilution
Serial dilution
Isotope dilution
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Serial dilution
Dilution
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Serial dilution
Dilution
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Sephadex
Colony-stimulating factor
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Summary and ConclusionsFresh rabies passage virus was diluted in tenfold serial dilutions up to 1:10,000,000 with nine different diluents. The four highest dilutions of each were tested in mouse groups of four, immediately on preparation and at varying intervals thereafter up to 24 hours. After 21 days the virus titers in mice were compared to determine the degree of unfavorable effect of each diluent on the virus.Results showed that the several diluents differed considerably in their harmful effects on the virus. Distilled water was slightly less harmful to the virus than hormone broth. Normal saline solution was definitely harmful in one hour. When a 10 per cent normal serum was added to a diluent its unfavorable effect on the virus was reduced. Serum tyrode solution was found to be the least harmful of the diluents tested and the most regular in its action. Serum water was only slightly less satisfactory and is recommended on account of the ease of its preparation.
Diluent
Serial dilution
Distilled water
Dilution
Physiological saline
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Luteinizing hormone (LH) potency estimates were made on unconcentrated aliquots from 10 urine pools by radioimmunoassay and on concentrated extracts from 8 of the same urine pools by both radioimmunoassay and bioassay. Urine specimens from groups of male adults and prepubertal children and several patients with disorders of sexual development were compared. The data from radioimmunoassay of unprocessed urine showed a very narrow range of differences in LH potency estimates between the different groups, whereas both radioimmunoassay and bioassay of processed urine revealed a much greater range of differences between the different groups of subjects. There was substantial agreement between LH potency estimates on extracted urine obtained by bioassay and radioimmunoassay. It is concluded that radioimmunoassay with its attendant advantages can be substituted for bioassay in the measurement of LH on urine extracts from patients excreting relatively small amounts of gonadotropin, thus facilitating studies of variation in LH excretion in physiologic and pathologic states.
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