LGC eBook - 2017 - 14

Accelerating Science Using Integrated PCR Tools * PCR Shows Off Its Clinical Chops

that can be as fast as 10 minutes from
blood sample to result. Their portable
devices for molecular diagnostics can
be used as genetic biosensors to bring
complex clinical testing directly to the
patient.
"Instead of sequential steps, reactions
happen in parallel, significantly
reducing analysis time. Buffer, enzyme,
and temperature profiles are optimized
to maximize sensitivity," explained
Sabrina Li, CEO, Coyote Bioscience.
"Both RNA and DNA can be analyzed
simultaneously from a drop of blood
in the same reaction."
The first-generation Mini-8 system
was used for Ebola detection in Africa,
where close to 600 samples were tested
with 98.8% sensitivity. Recently in
China, the Mini-8 system was applied
in hospitals and small community
clinics for hepatitis B and C, and Bunia
virus detection. The second-generation
14

| GENengnews.com

InstantGene system is currently being
tested internally with clinical samples.
Digital PCR
Personalized cancer care requires ultrasensitive detection and monitoring
of actionable mutations from patient
samples. The high sensitivity and precision of droplet-digital PCR (ddPCR)
from Bio-Rad Laboratories offers critical
advantages when clinical samples are
limited, degraded, or contain PCR
inhibitors.
Typically, formalin-fixed and paraffinembedded (FFPE) tissue samples are
processed. FFPE samples work well for
immunohistochemistry and protein
analysis; however, the formalin fixation
can damage nucleic acids and inhibit
the PCR reaction. Samples may yield
100 ng of purified nucleic acid, but the
actual amplifiable material is less than
1%, or 1 ng, in most cases.

Conventional real-time PCR technology, while suited to the
analysis of high-quality clinical samples, may effectively
conceal amplification efficiency changes when sample
quality is inconsistent. A more effective alternative, Bio-Rad
suggests, is its droplet-digital PCR (ddPCR) technology, which
can provide absolute quantification of target DNA or RNA,
a critical advantage when samples are limited, degraded, or
contain PCR inhibitors. The company says that of the halfdozen clinical trials that are using digital PCR, half rely on
the Bio-Rad QX200 ddPCR system.

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LGC eBook - 2017

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