FastPlex™ Norovirus multiplex One-Step digital PCR Detection Kit

Noroviruses belong to the family Caliciviridae. They are a group of non-enveloped, single-stranded, positive-sense RNA viruses that mainly cause acute gastroenteritis or gastrointestinal inflammation. Norovirus is characterized by rapid mutation, strong environmental resistance and low infectious dose. It is extremely adaptable to the environment and can survive in a variety of environments, providing favorable conditions for its spread. The virus is mainly transmitted through the fecal-oral route, aerosol transmission and contact. Symptoms of norovirus infection usually appear within 12 to 48 hours after exposure and include nausea, vomiting, abdominal pain, fever, headache and diarrhea.

Intended Use

This kit is designed for the detection of Norovirus GI and Norovirus GII nucleic acids. The results can be used as an aid in the diagnosis of patients with norovirus infection or suspected infection, and can also be used to detect norovirus nucleic acids in the environment and food.

The results of this kit are for diagnostic reference only and should not be used as the sole criterion for diagnosis. It is recommended to analyze the condition in conjunction with the patient's clinical manifestations and other laboratory test results.

Product Features

The detection methods for norovirus include electron microscopy, immunological methods, and molecular methods. Molecular methods include RT-qPCR, RT-LAMP, gene chip and so on. Among them, RT-qPCR  is known as the gold standard for detecting norovirus but has disadvantages such as the need to use a standard curve for quantification and the reaction being affected by inhibitors. Digital PCR has effectively solved this problem. The detection process does not require a standard curve and can perform absolute quantitative analysis of the virus load without relying on the Ct value. In addition, the unique nanoliter reaction system of digital PCR can reduce the impact of PCR inhibitors and is compatible with larger sample loading volumes. The multi-plex assay also improve the detection efficiency.

The advantages of digital PCR method in detection norovirus are as follows:

High precision : The FastPlex™ kit can provide absolute quantification of the target nucleic acid without the need for a standard curve. It can accurately determine the copy number of the norovirus, which is especially useful for the detection of low - abundance samples.

High resistance to inhibitors : The unique nanoliter reaction system of digital PCR can reduce the impact of PCR inhibitors. This is important for the detection of samples with complex matrices, such as environmental and food samples, where inhibitors may be present.

High sensitivity : The FastPlex™ kit can detect a small number of target molecules in a large background of non - target molecules. This makes it possible to detect norovirus in samples with low viral loads.

High reproducibility : The results of test are less affected by experimental conditions and operators, and have good reproducibility. This is important for the consistency and reliability of detection results.

Detection of multiple genotypes in a single sample: The FastPlex™ kit can simultaneously detect Norovirus GI and/or Norovirus GII. The detection probe for Norovirus GI is labeled with HEX fluorescence, the detection probe for Norovirus GII is labeled with FAM fluorescence.

Ø One  step reverse transcription assay .

Ø  It can detect various targets such as feces, sewage, food, patients, and suspected patients.

Ø  A single well can simultaneously differentiate different genotypes (Norovirus GI, Norovirus GII, control phage MS2, and human internal control).

Ø  Absolute quantification of the virus can be as low as a single copy.

Rainsure's one-step norovirus detection kit has strict quality control

(ROX channel for monitoring extraction efficiency)

	FAM （GⅡ）	HEX （GⅠ）	ROX （MS2）	Cy5 （Humna internal reference）
Positive Contro	-	-	-	-
Negative Control	+	+	+	+

The analysis of test results is simple.

No.	FAM （GⅡ）	HEX （GⅠ）	ROX （Extraction Control）	Cy5 （Human Internal reference）	Results
1	-	-	+	-	Negative
2	+	-	+/-	+/-	GII Positive
3	-	+	+/-	+/-	GI Positiv
4	+	+	+/-	+/-	GI&GIIPositive

The excellent linearity performance of FastPlex™ Norovirus multiplex One-Step digital PCR Detection Kit

Coparation of dPCR and qPCR method

	dPCR result		qPCR result			dPCR result		qPCR result
SampleName	HEX（GI） cps/ul	FAM（GII） cps/ul	Phenotype	Ct value	SampleName	HEX（GI） cps/ul	FAM（GII）  cps/ul	Phenotype	Ct value
NOV_1	0	6.14	GⅡ	28.84	NOV_32	1.86	0	GⅠ	21
NOV_2	0	3.48	GⅡ	22.58	NOV_33	17397	0	GⅠ	18.7
NOV_3	0	0.34	GⅡ	25.19	NOV_34	861	0	GⅠ	23.3
NOV_4	18145	0	GⅠ	30.86	NOV_35	224	0	GⅠ	25
NOV_5	28.7	24.6	GⅠ/GⅡ	24.33；22.83	NOV_36	14.3	0	GⅠ	22
NOV_6	247	0	GⅠ	21.25	NOV_37	12.8	0	GⅠ	24
NOV_16	0	253	GⅡ	16.07	NOV_38	21.3	0	GⅠ	25
NOV_18	0	0.207	GⅡ	16.73	NOV_39	0	439	GⅡ	22.64
NOV_20	0	2.26	GⅡ	30.19	NOV_40	0	179	GⅡ	23.91
NOV_21	0	4.54	GⅡ	17.43	NOV_42	0	582	GⅡ	22.24
NOV_22	0	6.62	GⅡ	30.13	NOV_43	0	35.4	GⅡ	24
NOV_23	1000000	859	GⅠ/GⅡ	19.55；23.54	NOV_44	0	1467	GⅡ	25
NOV_24	0	1000000	GⅡ	19.89	NOV_45	0	1.92	GⅡ	22.6
NOV_25	0	127	GⅡ	22.42	NOV_46	0	34.8	GⅡ	24.5
NOV_26	0	66.4	GⅡ	22.99	NOV_47	0	541	GⅡ	21.6
NOV_27	0	0.872	GⅡ	26.37	NOV_48	0	179	GⅡ	24
NOV_28	81.3	243	GⅠ/GⅡ	26.34；23.82	NOV_49	0	443	GⅡ	22
NOV_30	0.288	2.3	GⅠ/GⅡ	25.92；24.22	NOV_50	0	8599	GⅡ	20
NOV_31	146	0	GⅠ	24.7