Series: Ernestly measurement
Measurement of norovirus
You may have a sudden stomach ache in the middle winter.
You think of what you ate the day before, pour thinned hypochlorous acid in a spray bottle, and occupy the restroom for a long time.
According to the “2017 food poisoning report” issued by the Japanese Ministry of Health, Labour and Welfare, 20% of food poisoning was caused by norovirus when the cases are sorted by cause. It was the second leading cause with over 200 reported cases that year. Every year, the primary cause of food poisoning is either norovirus or campylobacter.
We examined norovirus using a mobile PCR device.
Synthesized RNA of Norovirus GII (ATCC: VR-3235SD)
THUNDERBIRD Probe One-step qRT-PCR Kit (TOYOBO)
|μL||Remarks (final concentration)|
|RT Enzyme Mix||0.5||0.5μL/reaction|
|2x Reaction Buffer||8.5||1x|
|COG2F forward primer||0.136||800nM|
|ALPF forward primer||0.136||800nM|
|COG2R reverse primer||0.136||800nM|
|RING1-TP(b) probe (FAM added)||0.255||150nM|
|PCR grade water||5|
Reaction sample total 17μL
In the THUNDERBIRD Probe One-step qRT-PCR Kit (TOYOBO) we used in the test, the RNA amount (/reaction) was detectable at the range of 4.4 x 105 to 4.4 x 10 copies, demonstrating that a correlation with the Ct value was maintained.
When electrophoresis was performed for 4.4 x 100 copies/reaction in the condition, almost no assumed PCR product was observed and any correlation with the Ct value was poor ⇒ almost no expected PCR product was observed and correlation with the Ct value was poor As a result, we determined that the results were not valid.
It is difficult to evaluate kits for RT made by various companies as conditions were different.
However, it can be said that excellent results were provided from a viewpoint of sensitivity in the conditions listed for the kit.