Glucoamylase-labeled nanogold flowers for in situ enhanced sensitivity of a glucometer-based enzyme immunoassay

2015 ◽  
Vol 7 (2) ◽  
pp. 507-512 ◽  
Author(s):  
Xiaohong Fu ◽  
Kun Xu ◽  
Jun Ye ◽  
Jie Chen ◽  
Xueyu Feng

Methods based on enzyme labels have been developed for glucometer-based immunoassays, but most involve low sensitivity and are unsuitable for routine use.

The Analyst ◽  
2020 ◽  
Vol 145 (12) ◽  
pp. 4164-4172 ◽  
Author(s):  
Li-Juan Zhi ◽  
Ai-Li Sun ◽  
Dianping Tang

Methods based on prussian blue nanoparticles (PBNPs) have been reported for photothermal immunoassays in analytical nanoscience fields but most suffer from low sensitivity and are not beneficial for routine use.


1995 ◽  
Vol 43 (8) ◽  
pp. 829-841 ◽  
Author(s):  
A Trembleau ◽  
F E Bloom

Although oligonucleotide probes are useful for in situ hybridization, their low sensitivity compared to riboprobes and cDNA remains a problem. We have systematically examined the protocols to provide a general procedure that increases the sensitivity of oligoprobes for light and electron in situ hybridizations by using mixtures of multiple non-overlapping oligonucleotides (multi-oligoprobes). The protocol achieves these improvements with both radioactive and non-radioactive oligoprobes. With 33P-labeled probes in a semiquantitative assay, we found that mixtures of up to six vasopressin-directed multi-oligoprobes, each employed at saturating concentration, led to an additive signal with no significant increase of the background. Using this approach with non-radioactive oligoprobes, we were able to detect in the hypothalamus several low or moderately abundant mRNAs, such as vasopressin heterogeneous nuclear RNA and the galanin, dynorphin, and tyrosine hydroxylase mRNAs. Moreover, we showed that multi-oligoprobes used in a pre-embedding procedure were suitable for studying the ultrastructural compartmentalization of moderately abundant mRNAs. Finally, with the same basic approach we demonstrated that two sets of multi-oligoprobes can be combined for simultaneous detection of two different mRNAs using fluorescent dyes, making this approach suitable for high-resolution confocal analyses. Overall, our data demonstrate that multi-oligoprobes provide a sensitive tool of choice for various applications in which both well-preserved morphology and high sensitivity are needed. In particular, these probes appear ideal for study of the comparative subcellular localization of mRNAs at both the light and the electron microscopic level.


1977 ◽  
Vol 30 (9) ◽  
pp. 894-898 ◽  
Author(s):  
J Kacaki ◽  
G Wolters ◽  
L Kuijpers ◽  
A Schuurs

2009 ◽  
Vol 12 (3) ◽  
pp. 195-199 ◽  
Author(s):  
Naim K. Fanaian ◽  
Cynthia Cohen ◽  
Sandra Waldrop ◽  
Jennifer Wang ◽  
Bahig M. Shehata

Detection of Epstein-Barr virus (EBV) may be achieved by various methods, including EBV-encoded RNA (EBER) in-situ hybridization (ISH) and immunohistochemistry (IHC) for latent membrane protein (LMP-1). We compared novel automated ISH and IHC techniques in pediatric lymphoproliferative disorders with results obtained by manual ISH. Thirty-seven pediatric cases previously studied by manual EBER ISH (including 18 EBER-positive, 15 EBER-negative, and 4 EBER-equivocal cases) were used for the study. Automated EBER ISH and automated LMP-1 IHC were performed using the BondMax autostainer and prediluted EBER probe and EBV cell surface 1 to 4 at 1:50 dilution, respectively. Results of each of the automated techniques for EBV detection were compared with results by manual EBER ISH. Compared with manual EBER ISH as the gold standard, automated ISH had a sensitivity and specificity of 94% and 69%, respectively, accuracy of 83%, positive predictive value (PPV) of 79%, and negative predictive value (NPV) of 90%. Automated IHC had a sensitivity of 44%, specificity of 93%, accuracy of 67%, PPV of 88%, and NPV of 59%. Automated ISH and IHC correlated significantly ( P < 0.045). Automated ISH is useful for diagnosis of EBV-related pediatric neoplasms, being easy to perform and interpret and requiring only the technologist's time to set up and having a high sensitivity and NPV. The automated IHC protocol is of too low sensitivity for routine use, although results show high specificity and PPV.


2015 ◽  
Vol 7 (5) ◽  
pp. 1843-1848 ◽  
Author(s):  
Sheng Wu ◽  
Jinsuo Chen ◽  
Yujian Tian ◽  
Xiaoming Tang ◽  
Wei Li ◽  
...  

A new glucometer-based enzyme immunoassay was developed for the sensitive detection of alpha-fetoprotein using biofunctional dendritic polyaniline nanofibers as the signal-transduction tag.


1981 ◽  
Vol 15 (1) ◽  
pp. 41-44 ◽  
Author(s):  
J. C. Cox ◽  
R. Horsburgh ◽  
D. Pye

Rabbit antibodies against Encephalitozoon cuniculi were detected in an enzyme immunoassay procedure in which antigen was grown and used in situ. The test appeared to be more sensitive than the indirect immunofluorescence test with which it was compared, but gave essentially the same results for the 64 sera evaluated. This procedure will allow any laboratory with cell-culture facilities to produce a diagnostic antigen without the need for antigen processing. It is simple and reliable, and does not require specialized equipment or microscopic assessment.


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