Overview
K.septempunctata has a wide host-specificity and infects various fish species such as salmon, flounder, mackerel and yellowtail. it is suspected to be a causative agent of foodborne illness in humans.
The novel K. septempunctata isolated from fish forms multiple white cyst structures or causes the dissolution of postmortem muscle, resulting in a phenomenon known as “Jelly meat”, which reduces the value of the product.
AccuPower® Kudoa septempunctata PCR Kit enables the detection of K. septempunctata, a type of myxosporean parasite, using PCR amplification with DNA extracted from fish muscle. Bioneer's patented enzyme-mediated HotStart technology is applied to suppress non-specific amplification and enhance reaction specificity and PCR efficiency. This product includes a positive control that aids in determining the presence of K. septempunctata.
K.septempunctata is a newly identified myxosporean parasite of Paralichthys olivaceus and a suspected causative agent of several food-borne gastroenteritis outbreaks. It parasitizes in fish's muscles then forms many white particulate cysts, with broad host specificity into salmon, halibut, mackerel, and tuna. It also causes the jelly meat phenomenon to dissolve the fish muscle after death, thereby reduces the value of the fish. It is possible to detect K. septempunctata by PCR using DNA extracted from fish muscle.
Features and Benefits
- Convenience
All necessary reactants for PCR are included in a tube (i.e., DNA polymerase, dNTPs, Buffer, Specific primers), allowing the users to perform reaction simply by adding template DNA and DEPC-D.W. No extra tracking dye is required for electrophoresis, which enables direct electrophoresis after the PCR reaction.
- Sensitivity
The differentiated PyroHotstart technology can effectively amplify trace amounts of template DNA by minimizing the generation of nonspecific amplification products and maximizing the PCR reaction efficiency.
- Stability
Contains a stabilizer in the PCR reaction mixture, making the PreMix type more stable than the solution type products.
- Reproducibility
The Bioneer mass production system follows the ISO 9001 quality system, providing minimized lot-to-lot variation and reproducible experimental results even in repeated experiments.
▶ Specification
| Components |
Amount |
|
AccuPower® Kudoa septempunctata PCR Premix
|
8-well strip x 12 ea |
Kudoa-KS Positive Control
(PC)DNA(1X105 copies/μl) |
105 μl x 2 ea |
| DEPC-D.W. |
1.5ml x 2 ea |
| Product manual |
1 ea |
▶ Experimental data
Figure 1. Evaluation of AccuPower® Kudoa septempunctata PCR Kit
Amplification of Kudoa-KS gene (357 bp) using AccuPower® Kudoa septempunctata PCR Kit. Sample was prepared by 10-fold serial dilution (5 x 105 copies/rxn ~ 50 copies/rxn). PCR was performed at 95℃ for 5 min, at 95℃ for 5 sec / at 60℃ for 30 sec / at 72℃ for 30 sec (40 cycles), and at 72℃ for 5 min.
Figure 2. Specificity of AccuPower® Kudoa septempunctata PCR Kit
The gDNA was extracted from the muscles of the Paralichthys olivaceus. PCR was performed at 95℃ for 5 min, 95℃ for 5 sec / 60℃ for 30 sec / 72℃ for 30 sec (40 cycle), and 72℃ for 5 min. Please note that the samples with PCR band (#2, 5 and 6) were also determined as clinically positive through microscopic examination method.
Figure 3. Reproducibility comparison between PCR cyclers
Sample was prepared by 10-fold serial dilution (5 x 105 copies / rxn to 5 x 102 copies / rxn). PCR was performed at 95℃ for 5 min, at 95℃ for 5 sec / at 60℃ for 30 sec / at 72℃ for 30 sec (40 cycles), and at 72℃ for 5 min. As a result of the analysis, it was confirmed that there was no significant difference among the equipment.
◈ Manual
• Manual_AccuPower® Kudoa septempunctata PCR Kit
◈ MSDS
• MSDS_AccuPower® Kudoa septempunctata PCR Kit
◈ Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.
• ISO 9001 - certificate
