PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings

Abstract

Molecular diagnostics are revolutionising the clinical practice of infectious disease. Their effects will be significant in acute-care settings where timely and accurate diagnostic tools are critical for patient treatment decisions and outcomes. PCR is the most well-developed molecular technique up to now, and has a wide range of already fulfilled, and potential, clinical applications, including specific or broad-spectrum pathogen detection, evaluation of emerging novel infections, surveillance, early detection of biothreat agents, and antimicrobial resistance profiling. PCR-based methods may also be cost effective relative to traditional testing procedures. Further advancement of technology is needed to improve automation, optimise detection sensitivity and specificity, and expand the capacity to detect multiple targets simultaneously (multiplexing). This review provides an up-to-date look at the general principles, diagnostic value, and limitations of the most current PCR-based platforms as they evolve from bench to bedside.

Virus-inducible reporter genes as a tool for detecting and quantifying influenza A virus replication

“…Although they have the potential to become standard methodology, some questions exist about the sensitivity and reliability of the tests (Landry and Ferguson, 2003;Ruest et al, 2003). Finally, nucleic acidbased techniques such as real time multiplex PCR (Ellis and Zambon, 2002;Yang and Rothman, 2004) or microarrays (Kessler et al, 2004;Wang et al, 2002) can provide specific, sensitive and rapid diagnosis of clinical samples but are expensive and require significant expertise.…”

Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis

“…Appropriate care and correct conditioning of the sample may prevent such problems, but the integrity of target DNA must be monitored. Methods that ensure the best sample processing include incorporation of internal amplification controls into the PCR assay to monitor the presence of purified sample DNA and potential PCR inhibitors (40).…”

Comparison of real-time PCR and conventional PCR for detection of Leishmania (Leishmania) infantum infection: a mini-review

“…The methodology requires, first, the identification, at least in part, of the DNA target for the development of primers or probes that will hybridize specifically to the target sequence. With the increased number of sequenced pathogen genomes, catalogs of DNA sequences can be exploited for the development of diagnostic tests based on PCR, as a result, since the past decade, numerous tests commercially available are based on this technique (7).…”

“…In the past two decades, cnPCR technique has been modified to expand its use and versatility (7). The possibility of using it in the same reaction, on a pair of primers with simultaneous amplification for multiple target DNA sequences is called multiplex-PCR.…”