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[Exploration and analysis of PCR-inhibitory components in peripheral blood and bone marrow samples]
Ke-Feng Shen1, Mo Yang1, Qian-Li Jiang1
1Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China.
Abstract:
PCR is an enabling and preferred technology for the detection of DNA or mRNA in genomic age, and has been widely used in molecular medicine, biotechnology, microbiology and diagnostics. PCR has many excellent traits, such as high specificity and sensitivity, short handle time, reliable and effective results, low demand for sample. In recent years, real-time reverse transcription PCR (qRT-PCR) has acquired an impressive improvement in the detection of mRNA, it has a perfect sensitivity and can quantify some rare mRNA copies as low as less ten. The progress in PCR results in a more strict demand for sample, so it is necessary to take the potential PCR-inhibitory components in peripheral blood and bone marrow samples into account again. This review explores the potential PCR-inhibitory components in peripheral blood and bone marrow samples, and discusses the possible mechanism and solutions for the inhibition effect, also emphasizes the potent effect of the frequently-used anticoagulant heparin sodium in PCR. This article will be helpful to assess the PCR detection results (especially qRT-PCR ) in some degree, for example, can use it to decide whether the PCR detection results are reasonable for some low-expression genes, or use it to eliminate the false negative results generated by PCR-inhibitory components in sample.
Insights
Polymerase Chain Reaction (PCR) is crucial for DNA/mRNA detection. This review examines inhibitors in blood samples, like heparin, that can affect PCR results, especially for low-expression genes.
Area of Science:
- Molecular biology
- Genomics
- Biotechnology
Context:
- Polymerase Chain Reaction (PCR) is a cornerstone technology in molecular biology, widely applied in medicine, biotechnology, and diagnostics for DNA and mRNA detection.
- Real-time reverse transcription PCR (qRT-PCR) has significantly advanced mRNA detection sensitivity, enabling quantification of rare transcripts.
- Increased PCR sensitivity necessitates a re-evaluation of potential inhibitory substances in biological samples, particularly peripheral blood and bone marrow.
Purpose:
- To explore potential PCR-inhibitory components present in peripheral blood and bone marrow samples.
- To discuss the mechanisms underlying PCR inhibition and propose solutions.
- To highlight the significant inhibitory effect of heparin sodium, a common anticoagulant, on PCR assays.
Summary:
- This review identifies and analyzes PCR inhibitors found in blood and bone marrow, focusing on their mechanisms and mitigation strategies.
- Heparin sodium is identified as a potent inhibitor that can compromise PCR and qRT-PCR accuracy.
- Understanding these inhibitors is crucial for accurate interpretation of PCR results, especially for low-expression targets.
Impact:
- Provides guidance for assessing the reliability of PCR and qRT-PCR results, particularly in clinical diagnostics.
- Helps in troubleshooting false-negative results caused by PCR inhibitors in blood-derived samples.
- Enhances the accuracy of molecular diagnostic assays by accounting for sample-specific inhibition effects.
