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

Abstract

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.

Detection of respiratory syncytial virus using nanoparticle amplified immuno-polymerase chain reaction

“…Even fragments of cell wall that have the protein of interest expressed can be detected. Secondly, each virion may contain only a single copy of RSV genomic RNA, but the protein targets are often expressed at a higher copy number, each one of which can act as a potential target [26]. …”

“…The original I-PCR design has been modified in a number of ways to strengthen various aspects of the assay. Different antibody-DNA linking strategies have been used; from streptavidin-protein A coupling, to strictly streptavidin-biotin coupling, and even phage display mediated I-PCR (PD-IPCR), where a single chain variable fragments are displayed on the phage and phage DNA itself acts as a DNA tag [27], [28], [26] and [29]. Capture antibodies have been bound to biogenic magnetosome nanoparticles (M-IPCR) instead of protein binding plates to enhance signal generation [30].…”