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. 2016 Nov 11;23(6):444–463. doi: 10.1111/xen.12277

Figure 1.

Figure 1

Viral Screening of Porcine Islets by qPCR / qRT‐PCR. Overview of the qPCR/qRT‐PCR (as appropriate) testing strategy used in the viral screening of porcine islet macrobeads (Checkpoint 3). 1st Step: each test sample is spiked with an internal extraction control (IEC; plant RNA or DNA) at the detection limit, mimicking the target nucleic acid. This process allows the level of nucleic acid recovery in the presence of the test sample to be assessed, which ensures that low levels of RNA or DNA can be recovered. 2nd Step: Test samples contain viral target primers/probes, multiplexed with internal positive control (IPC) reagents during amplification. In a separate assay, test samples are spiked with target nucleic acid (viral RNA or DNA) at the detection limit. This process ensures any negative results are not caused by sample matrix interference of the qPCR assay. This method of viral screening complies with the specific testing approach using PCR detection as described in Ph. Eur. 2.6.21, USP <1237>, and FDA Vaccine Guidance to Industry (2010)