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Variola virus (VARV), the causative agent of smallpox, is estimated to have killed between 300 million and 500 million people in the 20th century and was responsible for widespread mortality and suffering for at least several preceding centuries. Humans are the only known host of VARV, and smallpox was declared eradicated in 1980. The timeline of the emergence of smallpox in humans is unclear.
A real-time polymerase chain reaction (real-time PCR,or qPCR) is a laboratory technique of molecular biology based on the polymerase chain reaction(PCR). It monitors the amplification of a targeted DNA molecule during the PCR(i.e.,in real time), not at its end, as in conventional PCR.
Two common methods for the detection of PCR products in real-time PCR are(1)non-specific fluorescent dyes that intercalate with any double-stranded DNA and (2)sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter, which permits detection only after hybridization of the probe with its complementary sequence.