Direct RT-PCR Kit is designed for the amplification and detection of target RNA directly from whole blood, animal tissues, and plant tissues without any RNA purification processes. This kit contains enzyme mixture including HelixCript™ Thermo Reverse Transcriptase and antibody-coupled Taq polymerase, and 2x Buffer mix containing dNTPs, MgCl₂, and unique buffer system to resist various PCR inhibitors from tissue samples. A Uracil-DNA glycosylase and dUTP applied version is also available.

 

Figure 1. Comarison of multiple detection for swine influenza virus by direct RT-PCR with conventional RT-PCR from cultured virus. Lysate of influenza viral particles were applied into direct RT-PCR for multiple detection. Efficiency of detection sensitivity for direct RT-PCR was compared with conventional RT-PCR using total RNAs purified from the same amount of viral particles. NTC: Negative control.

Figure 2. Comparison of amplification efficiency of direct RT-PCR with conventional RT-PCR from whole blood(A) or buccal swap(B). Lysate of each sample were used for direct RT-PCR as indicated volume above. For the comparison with reaction, the same amount of each sample were used in total RNA purification and the purified total RNAs were applied in conventional RT-PCR as indicated volume above. NTC: Negative control.

Figure 3. Comparison of detection sensitivity of viral target in direct RT-PCR with conventional RT-PCR from animal RNA virus-spiked whole blood. Lysate of blood spiked with virus particles were used for direct RT-PCR. Total RNAs purified from the same amount of virus-spiked blood were used for conventional RT-PCR. NTC: Negative control.

Figure 4. Comparison of detection sensitivity of direct RT-PCR with conventional RT-PCR from Virus-infected plant seeds. Melon necrotic spot virus(MNSV)-infected melon seeds were used in this experiment. After mixing a grain of crushed seed with dilution buffer, lysate were used for direct RT-PCR. Total RNAs purified from the same amount of MNSV-infected plant seed were used for conventional RT-PCR. NTC: Negative control.

Application 

• Point-of -Care molecular diagnostics
• Direct amplification of target RNA from samples
• Direct detection of RNA viral pathogens from various tissues 

Quality control

Functional Assay : HelixAmp™ Direct RT-PCR Kit is evaluated by amplification of RT-PCR product corresponding to each sample type-specific RNA region directly using whole blood, plant leaf tissue, or animal tissue according to protocol.