Enhanced luminescence procedure for sensitive determination of peroxidase-labelled conjugates in immunoassay

Whitehead, T. P.; Thorpe, G. H. G.; Carter, Timothy J. N.; Groucutt, Carol; Kricka, Larry J.

doi:10.1038/305158a0

Cited by 340 publications

(106 citation statements)

References 7 publications

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“…The amplified products of T. cruzi DNA detected in samples from babies 1, 3, and 4, by agarose gel electrophoresis and Southern blot hybridization analysis 21 have been previously reported. 19 It was possible to detect T. cruzi DNA in the plasma of six newborns on the day of birth.…”

Section: Distribution Of Infectedmentioning

confidence: 99%

Treatment of congenital Chagas' disease diagnosed and followed up by the polymerase chain reaction.

Russomando¹,

Tomassone²,

Guillén³

et al. 1998

The American Journal of Tropical Medicine and Hygiene

134

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Abstract. In 1991 and 1992, a prenatal screening of Trypanosoma cruzi infection was carried out using ELISA and indirect immunofluorescence techniques. A total of 840 blood samples from pregnant women, obtained at the Maternity Ward of the Hospital de Clínicas, National University of Asunción (Asunción, Paraguay), and 1,022 samples from the Regional Hospital of the San Pedro Department of Paraguay were examined. It was observed that 7.7% and 10.5%, respectively, of the pregnant women were serologically positive for infection with T. cruzi. When blood samples obtained from newborns on the day of birth or, at the most, on the first few days afterwards were examined by direct microscopic observation, an incidence of congenital transmission of 3% was found. These results are consistent with those of neighboring countries. When a serologic follow-up was conducted on the newborns until six months of age, the incidence of congenital transmission reached 10%. The same incidence rate was obtained when the samples collected during the first days after birth were examined by the polymerase chain reaction (PCR). Fiftyeight infants born to seropositive mothers were followed-up, two of which were positive by direct microscopic observation at birth, and four who were PCR-positive, but microscopy-negative at birth. None of the infants were positive for IgM at birth. The infected babies were treated with benznidazole and were followed-up by serology and PCR for four years. We conclude that the PCR has a clear advantage over conventional techniques for the early detection of congenital transmission of T. cruzi infection, and for monitoring infants undergoing chemotherapy.

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Section: Distribution Of Infectedmentioning

confidence: 99%

Treatment of congenital Chagas' disease diagnosed and followed up by the polymerase chain reaction.

Russomando¹,

Tomassone²,

Guillén³

et al. 1998

The American Journal of Tropical Medicine and Hygiene

134

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“…Thus quantitative HBsAg can be a promising prognostic marker of the natural history of HBV infection as well as during antiviral therapy. The first quantitative assays measuring HBsAg levels using enhanced chemiluminescence were introduced over 20 years ago (43,44) but were limited by a lack of appropriate standardization. Initially, HBsAg levels were expressed as serial dilutions of a reference sample from the Paul Ehrlich Institute (Langen, Germany) (44).…”

Section: Hbsag Revisitedmentioning

confidence: 99%

Quantification of hepatitis B surface antigen: a new concept for the management of chronic hepatitis B

Moucari

Marcellin

2011

Liver International

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HBsAg is a very important clinical test that might not only indicate active hepatitis B virus (HBV) infection but might also be used to predict clinical and treatment outcome. Clearance of HBsAg in patients with chronic HBV infection is associated with a much better clinical outcome, although surveillance for early detection of hepatocellular carcinoma (HCC) should continue. HBV DNA quantification is currently used for selecting candidates for therapy, monitoring response to therapy and detecting the emergence of drug resistance. Assays for HBsAg quantification are less expensive than HBV DNA and fully automated with a high throughput capacity. HBsAg titering may be a useful tool to manage patients with chronic HBV, to more clearly define which patients may, and more importantly, may not, benefit from treatment. Baseline and on-treatment HBsAg quantification may help to refine future treatment algorithms for both immune-modulator therapy and nucleos(t)ide analogues. Both HBV markers provide complementary information on the status of HBV infection. However, the relevance of serum HBsAg levels and its use as a reliable replacement for both covalently closed circular DNA and HBV DNA remain unclear.

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“…Briefly, proteins were extracted in 43 SDS sample buffer, fractionated in 10% SDS-PAGE mini gels, blotted to nitrocellulose membranes, stained with Ponceau S to verify equal loading, washed, probed with appropriate primary antibodies and the secondary antibody conjugated with horseradish peroxidase, and the signals developed using the enhanced chemiluminescence method (Whitehead et al, 1983). The SDS-PAGE running time, voltage, and temperature appear to determine the fine resolution associated with migration alterations resulting from protein phosphorylation.…”

Section: Immunostaining Immunoblot and Phosphorylation Analysismentioning

confidence: 99%

ArabidopsisCryptochrome 2 Completes Its Posttranslational Life Cycle in the Nucleus

et al. 2007

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CRY2 is a blue light receptor regulating light inhibition of hypocotyl elongation and photoperiodic flowering in Arabidopsis thaliana. The CRY2 protein is found primarily in the nucleus, and it is known to undergo blue light-dependent phosphorylation and degradation. However, the subcellular location where CRY2 exerts its function or undergoes blue light-dependent phosphorylation and degradation remains unclear. In this study, we analyzed the function and regulation of conditionally nuclear-localized CRY2. Our results show that CRY2 mediates blue light inhibition of hypocotyl elongation and photoperiodic promotion of floral initiation in the nucleus. Consistent with this result and a hypothesis that blue light-dependent phosphorylation is associated with CRY2 function, we demonstrate that CRY2 undergoes blue light-dependent phosphorylation in the nucleus. CRY2 phosphorylation is required for blue light-dependent CRY2 degradation, but only a limited quantity of CRY2 is phosphorylated at any given moment in seedlings exposed to blue light, which explains why continuous blue light illumination is required for CRY2 degradation. Finally, we showed that CRY2 is ubiquitinated in response to blue light and that ubiquitinated CRY2 is degraded by the 26S proteasome in the nucleus. These findings demonstrate that a photoreceptor can complete its posttranslational life cycle (from protein modification, to function, to degradation) inside the nucleus.

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Enhanced luminescence procedure for sensitive determination of peroxidase-labelled conjugates in immunoassay

Cited by 340 publications

References 7 publications

Treatment of congenital Chagas' disease diagnosed and followed up by the polymerase chain reaction.

Treatment of congenital Chagas' disease diagnosed and followed up by the polymerase chain reaction.

Quantification of hepatitis B surface antigen: a new concept for the management of chronic hepatitis B

ArabidopsisCryptochrome 2 Completes Its Posttranslational Life Cycle in the Nucleus

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