In-situ immuno-PCR to detect antigens

Cao, Yi; Kopplow, Kathrin; Liu, Guang-Yaw

doi:10.1016/s0140-6736(00)02696-9

Cited by 24 publications

(12 citation statements)

References 5 publications

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“…Specific examples of these infectious agents which have been analyzed using IPCR are Angiostrongylus cantonensis Ag of fifth-stage circulating worms (AcL 5 ) [48] , norovirus [25] , rotavirus VP6 Ag [24] , mumps-specific IgG [49,50] , hepatitis B surface Ag (HBsAg) [4,5,8,51] and Bacteriodes fragilis lipopolysaccharide (LPS) [52] .…”

Section: Detection Of Pathogens That Are Diffi Cult To Detect Purifymentioning

confidence: 99%

“…Cao et al were the first to report the in situ modification of the IPCR method to detect HBV in tissues [51] . A total of 17 explanted livers from HBV infected patients and 2 negative donor livers were analyzed for HBsAg expression.…”

Section: Barletta and Bartolomementioning

confidence: 99%

“…Similar to the method used by Cao et al [51] , Zhang et al applied in situ IPCR for the detection of a specific B. fragilis LPS in a mixture of microorganisms [52] . The LOD was 10 4 colony-forming units (cfu)/ml of B. fragilis in a bacterial suspension.…”

Section: Bacteroides Fragilis Lipopolysaccharidementioning

confidence: 99%

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Immuno-polymerase chain reaction as a unique molecular tool for detection of infectious agents

Barletta

Bartolome

2007

Expert Opinion on Medical Diagnostics

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Theoretically, the immuno-polymerase chain reaction (IPCR) method is the most sensitive technique for the detection of proteins and gains its uniqueness through the exponential amplification of a signal-generating nucleic acid intermediate attached to a protein target. This method is similar to PCR for the detection of nucleic acid targets, and has now been shown to offer the ability to detect infectious agents where nucleic acids are not present. Although the technical development of IPCR has taken a torturous path down a winding avenue of encouraging advances, the method remains rarely utilized by the scientific community and completely unused as a clinical diagnostic test approved by a national accrediting agency. Although the use of real-time instrumentation has enhanced the performance of IPCR to higher levels of statistical accuracy and reproducibility, as compared with the conventional method, its application remains limited by the high standards required for clinical diagnoses of infectious diseases. This review summarizes experimental data published to date describing the utilization of the IPCR method as it relates to the detection and diagnosis of human infectious disease, and examines the progressive development of this method, as well as the factors impeding its universal application as a clinical diagnostic tool. With further standardization and validation, the IPCR method has the potential to become the most analytically sensitive method available for the detection of target proteins of infectious diseases.

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Section: Detection Of Pathogens That Are Diffi Cult To Detect Purifymentioning

confidence: 99%

Section: Barletta and Bartolomementioning

confidence: 99%

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Immuno-polymerase chain reaction as a unique molecular tool for detection of infectious agents

Barletta

Bartolome

2007

Expert Opinion on Medical Diagnostics

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“…Also, immuno-PCR is capable to detect minimal amount of biological macromolecules, i.e. proteins, carbohydrates, and lipids, which is very useful for the clinical diagnosis and biological research (Cao et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Immuno-PCR in cancer and non-cancer related diseases: a review

Assumpção

Silva

2016

Veterinary Quarterly

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Polymerase chain reaction-amplified immunoassay (immuno-PCR, iPCR) is a method that combines the specificity of an immunological detection method and the sensitivity of a nucleic acid amplification method. In this way, immuno-PCR uses a minimum amount of sample, and allows the detection of rare diseases and those diseases in very early stage (i.e. infectious diseases, degenerative disorders, or neoplastic diseases). The present review was aimed to describe this new methodology and applications to the early detection of cancer and noncancer related diseases, and discuss about the possibility to detect diverse biomarkers of oncology disorders, such as breast, gastric, colorectal and nasopharynx cancer, and other factors related to the growth of the neoplastic disease.

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“…The signal, however, cannot be increased by increasing the reaction time on account of the short life of the tyramide radical, a key reactant in TSA (7). The more recently developed in situ immuno-PCR is more sensitive than TSA (8). Unfortunately, it has a problem inherent in in situ PCR (9); namely the dissociation and diffusion of labeled DNAs from the target protein.…”

Section: Introductionmentioning

confidence: 99%

Oligo(dA-dT)-dependent signal amplification for the detection of proteins in cells

et al. 2004

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An ultrasensitive protein detection system in situ named the ImmunoAT-tailing method was developed. It consists of three elementary processes: (i) detection of a protein by a primary antibody and a biotinylated secondary antibody; (ii) linking of biotinylated 15-base oligo(dA-dT) to the biotinylated immunocomplex via streptavidin; and (iii) self-priming elongation of oligo(dA-dT) by the Klenow fragment, 3' to 5' exo-. After the elongation reaction in the presence of dATP, dTTP, and dye-labeled dUTP, the protein was labeled with a large number of the dye molecules. The poly(dA-dT) elongated without the labeled nucleotides was detected by 4',6-diamidino-2-phenylindole (DAPI) staining. By combining the different labelings, double staining was possible. This ImmunoAT-tailing method has a specificity and sensitivity higher than that of tyramide signal amplification.

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In-situ immuno-PCR to detect antigens

Cited by 24 publications

References 5 publications

Immuno-polymerase chain reaction as a unique molecular tool for detection of infectious agents

Immuno-polymerase chain reaction as a unique molecular tool for detection of infectious agents

Immuno-PCR in cancer and non-cancer related diseases: a review

Oligo(dA-dT)-dependent signal amplification for the detection of proteins in cells

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