Time-resolved Fluorescence Immunoassay

2019 ◽  
pp. 2312-2313
Author(s):  
W. Stöcker ◽  
C. Krüger
Keyword(s):  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved

Development of a time-resolved fluorescence immunoassay based on immunomagnetic beads for gastrin-17

2021 ◽  
pp. 113179
Author(s):  
Shaoxiong Zheng ◽  
Renjing Hu ◽  
Xiaomei Yu ◽  
Lingli Chen ◽  
BinrongWang ◽  
...  
Keyword(s):  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Immunomagnetic Beads ◽  
Time Resolved

Establishment and clinical application of a highly sensitive TAT-2 time-resolved fluorescence immunoassay detection

2021 ◽  
Author(s):  
Xindong Chen ◽  
Jianfeng Hong ◽  
Han Zhao ◽  
Zhongyi Xiang ◽  
Yuan Qin ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Nasopharyngeal Cancer ◽  
Reaction Rates ◽  
Measurement Range ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Combined Use ◽  
Highly Sensitive ◽  
Positive Rate

Abstract Background: A rapid and highly sensitive assay for tumor-associated trypsinogen-2 (TAT-2) based on the time-resolved fluorescence immunoassay (TRFIA) detection technique was developed for the determination of serum TAT-2 levels in cancers. Results: The measurement range of TAT-2-TRFIA was 1.53-300 ng/mL. The within-run and between-run coefficients of variation of TAT-2-TRFIA were 4.38% and 7.82%, respectively. The recovery rate of TAT-2-TRFIA was 103.0%. The cross-reaction rates of trypsin and T-cell immunoglobulin mucin 3 were 0.02% and 0.82%, respectively. The TAT-2-positive rates in lung cancer, liver cancer, nasopharyngeal cancer, cholangiocarcinoma, brain cancer, and pancreatic cancer were 45.9%, 50.0%, 45.0%, 64.3%, 50.0%, and 41.7%, respectively, with the areas under ROC curves of 0.788, 0.734, 0.862, 0.720, 0.887, and 0.585, respectively. In patients with lung cancer, the positive rate of the single indicator CEA was 28.4%, which increased to 60.6% after combined use with TAT-2. In patients with cholangiocarcinoma, the positive rate of CA-199 was 35.7%, which increased to 71.4% after combined use with TAT-2. Conclusions: TAT-2 is expected to be used as an auxiliary diagnostic indicator for the combined use of tumor markers to improve the positive rate and accuracy of detection.

118 DEVELOPMENT AND EVALUATION OF A TIME-RESOLVED FLUORESCENCE IMMUNOASSAY FOR ESTRONE-1-SULFATE IN URINE AS A TOOL FOR DIAGNOSIS OF EARLY PREGNANCY IN SWINE

2008 ◽  
Vol 20 (1) ◽  
pp. 139
Author(s):  
Y. S. Park ◽  
S. H. Yang ◽  
S. M. Park ◽  
S. J. Kim ◽  
J. B. Kim
Keyword(s):  
Early Pregnancy ◽  
Field Trial ◽  
Dynamic Range ◽  
Sulfate Concentration ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Limit Of Quantification ◽  
Time Resolved ◽  
Pregnancy Diagnosis ◽  
Routine Basis

Early identification of pregnancy or non-pregnancy in sows is considered very important, as the management of sows during the post service period is crucial if the breeding efficiency of a herd is to be maximized. Studies of steroid hormones in pregnant sows showed that there was a significant increase in plasma estrone-1-sulfate concentration by the 16th day of gestation, which reaches peak values between Days 23 and 30 of gestation. Since plasma estrone-1-sulfate concentrations are high between Days 23 and 30 of pregnancy, its determination has been used as a means for early pregnancy diagnosis and monitoring fertility in sows. However, the application of the method in pig farms on a routine basis remains restricted because blood sampling is difficult and disturbs the animals. The present study describes the development of a simple and reliable time-resolved fluorescence immunoassay (TR-FIA) method for the estimation of estrone-1-sulfate in swine urine, which was assessed as a means for early diagnosis of pregnancy and monitoring fertility in sows. We demonstrated cross activity between Anti-estrone-1-glucuronide antibody (Clone 155) and estrone-1-sulfate. The method is based on a direct competitive heterogeneous immunoassay by the typical procedure of competitive immunocomplex formation. For detection of estrone-1-sulfate, anti-estrone-1-glucuronide antibody (Clone 155) was first coated on polystyrene microplates, and estrone-1-sulfate was captured by the primary antibody with estrone-1-glucuronide labeled with europium. The immunocomplex was subsequently dissociated by the enhancement solution containing Triton X-100, acetic acid, and chelators. The free europium was detection by DELFIA 1420 detector (Perkin-Elmer Life Sciences, Waltham, MA, USA). The fluorescence intensity of free europium at 613 nm was proportional to the logarithm of the concentration of estron-1-sulfate in a dynamic range of 0.078~10 ng mL–1. Intra-assay variation for estrone-1-sulfate was 4%. The limit of quantification was 100 pg mL–1. The mean estrone-1-sulfate concentration was significantly higher in pregnant sows (15.6 � 5.3 ng mL–1) than in non-pregnant sows and in sows in estrus (0.74 � 0.44 ng mL–1). Taking the concentration of 20 pg mL–1 as a cut-off, all cases of non-pregnant sows and sows in estrus were negative. Urine estrone-1-sulfate concentrations in pregnant sows at 23-day intervals post-service were 14~16 ng mL–1. According to the results of our field trial, urine estrone-1-sulfate concentrations are very low during estrus and remain low in non-pregnant sows at different stages of the estrous cycle, whereas the concentration increases significantly during specific stages of pregnancy at 23-day intervals. It is concluded that the satisfactory sensitivity of the present assay in combination with the good correlation for pregnancy from the present field trial makes this method a very useful technique for early pregnancy diagnosis in swine; the simplicity of urine sampling makes also it suitable for practical use.

Measurement of estrone-3-glucuronide in urine by rapid, homogeneous time-resolved fluoroimmunoassay.

1989 ◽  
Vol 35 (4) ◽  
pp. 555-559 ◽  
Author(s):  
G Barnard ◽  
F Kohen ◽  
H Mikola ◽  
T Lövgren
Keyword(s):  
Monoclonal Antibodies ◽  
Liquid Phase ◽  
Clinical Utility ◽  
Ovarian Function ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Europium Chelate ◽  
Covalently Linked ◽  
Sensitivity Specificity

Abstract We describe a liquid-phase nonseparation time-resolved fluorescence immunoassay for measuring estrone-3-glucuronide in undiluted urine. The sensitivity, specificity, and accuracy are similar to those for a conventional separation fluoroimmunoassay or radioimmunoassay, but the speed, convenience, precision, reliability, and clinical utility of the new method are more advantageous. The labeled antigen, a fluorescent europium chelate covalently linked to estrone-3-glucuronide, is incubated for 10 min with a limited concentration of polyclonal or monoclonal antibodies to estrone-3-glucuronyl-6-bovine serum albumin and 10 microL of standard or sample (undiluted urine) in microtiter wells. The fluorescence emanating from the antibody-free label, which is proportional to the concentration of estrone-3-glucuronide in the standard or sample, is then measured in a time-resolved fluorometer. The method is useful for monitoring ovarian function in women.

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