Sensitive PCR detection of viral nucleic acids plays a critical role in infectious disease research, diagnosis and monitoring. In the context of SARS-CoV-2 detection, recent reports indicate that digital PCR-based tests are significantly more sensitive than traditional qPCR tests. Numerous factors can influence digital PCR reaction sensitivity. In this review, using a model for human HIV infection and the Raindance ddPCR platform as an example, we describe technical aspects that contribute to sensitive viral signal detection in DNA and RNA from tissue samples, which often harbor viral reservoirs and serve as better predictors of disease outcome and indicators of treatment efficacy.
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【저자키워드】 HIV, Infectious diseases, COVID-19, Coronavirus disease 2019, SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2, HIV, Human immunodeficiency virus, virus, RNA, digital PCR, DNA, RSV, respiratory syncytial virus, SARS, Severe acute respiratory syndrome, Cure research, CT, Computed tomography, PIV, parainfluenza virus, SNP, single nucleotide polymorphism, LoD, limit of detection, NHP, Non-human primate, ddPCR, droplet digital polymerase chain reaction, HPV, Human Papillomavirus, MiRNA, MicroRNA, cART, combination antiretroviral therapy, Ct, threshold cycle, dMIQE, Minimum Information for Publication of Quantitative digital PCR experiments, dPCR, digital PCR, FFPE, formalin-fixed paraffin-embedded, HO, high occupancy, HTLV, human T-cell leukaemia virus, MGB, minor groove binder, NGS, next generation sequencing, qPCR, quantitative (real time) polymerase chain reaction, RT, reverse transcriptase, RT-dPCR, reverse transcription digital polymerase chain reaction, RT-qPCR, quantitative reverse transcription (real time) polymerase chain reaction, SIV, simian immunodeficiency virus, SNV, single nucleotide variant, STLV-1, simian T cell lymphotropic viruses type 1, STLV-2, simian T cell lymphotropic viruses type 2, Tm, melting temperature, PCR sensitivity,
【저자키워드】 HIV, Infectious diseases, COVID-19, Coronavirus disease 2019, SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2, HIV, Human immunodeficiency virus, virus, RNA, digital PCR, DNA, RSV, respiratory syncytial virus, SARS, Severe acute respiratory syndrome, Cure research, CT, Computed tomography, PIV, parainfluenza virus, SNP, single nucleotide polymorphism, LoD, limit of detection, NHP, Non-human primate, ddPCR, droplet digital polymerase chain reaction, HPV, Human Papillomavirus, MiRNA, MicroRNA, cART, combination antiretroviral therapy, Ct, threshold cycle, dMIQE, Minimum Information for Publication of Quantitative digital PCR experiments, dPCR, digital PCR, FFPE, formalin-fixed paraffin-embedded, HO, high occupancy, HTLV, human T-cell leukaemia virus, MGB, minor groove binder, NGS, next generation sequencing, qPCR, quantitative (real time) polymerase chain reaction, RT, reverse transcriptase, RT-dPCR, reverse transcription digital polymerase chain reaction, RT-qPCR, quantitative reverse transcription (real time) polymerase chain reaction, SIV, simian immunodeficiency virus, SNV, single nucleotide variant, STLV-1, simian T cell lymphotropic viruses type 1, STLV-2, simian T cell lymphotropic viruses type 2, Tm, melting temperature, PCR sensitivity,