Peste des petits ruminants virus (PPRV) nucleic acid test kit (Fluorescent RT-PCR) PPR ONE-STEP RTPCR KIT

Peste des petits ruminants virus (PPRV) nucleic acid test kit

(Fluorescent RT-PCR)

[Product Overview]

This product is a one-step qRT-PCR nucleic acid test kit designed for the auxiliary detection of PPRV nucleic acid in samples. Specific primers and a specific fluorescent probe targeting the conserved region of the PPRV N gene have been developed; the probe is labeled with the FAM fluorescent dye. When combined with the qRT-PCR reaction mix, this kit enables rapid detection of PPRV nucleic acid using a fluorescent quantitative PCR instrument.

The kit includes a positive control and a negative control. The positive control consists of a plasmid containing the amplification fragment, used to assess the validity of the assay; the negative control is used to detect the presence of contamination in the reaction system.

[Product Composition]

[Storage Conditions]

Store at-20 °C, protected from light; shelf life: 12 months.

[Applicable Instruments]

Real-time fluorescent PCR instruments, such as the ABI 7500, Bio-Rad CFX96, and Bio-Rad 96P.

[Sample Requirements]

• Customers are required to provide their own reagents: normal saline.
• Collected tissue samples—including nasal swabs, oral swabs, ocular swabs, as well as lymph nodes, spleen, kidney, rectum, colon, and lung—should be stored and transported at 2–8 °C and promptly submitted for laboratory testing. Alternatively, the samples may be preserved in viral transport medium and then sent to the laboratory for analysis.

Note: Samples submitted for testing must be fresh; repeated freeze-thaw cycles are strictly prohibited.

• It is recommended to use a commercial viral nucleic acid extraction kit for RNA extraction and purification; the extracted RNA can then be directly used for detection. If samples are not analyzed immediately after extraction, they may be stored at-80 °C for later use; however, repeated freeze-thaw cycles should be avoided.

[Interpretation of result]

1. Experimental setup requirements

The reported fluorescence for PPRV in this product is FAM.

1). Positive control: The FAM channel exhibited a characteristic S-shaped amplification curve, and all Ct values were ≤35;

2). Negative control: No Ct value detected in the FAM channel;

The above requirements must be met simultaneously within the same experiment; otherwise, the experiment will be invalidated, and it must be repeated.

• Judgment criteria

For suspicious results, a retest is required; if the Ct value in the retest is ≤45, the result is classified as positive; if no Ct value is obtained, the result is classified as negative.

[Additional notes]

• The target sequence detected by this assay kit is a conserved region of PPRV. However, if the virus undergoes a genetic mutation at this target sequence, false-negative results may occur.
• Test results depend on the collection, storage, and transportation conditions of the samples; errors in any of these stages can lead to false-negative results; conversely, cross-contamination during sample handling may result in false-positive results.
• This product is intended for research research use only. Before use, please carefully read this kit instruction manual and strictly follow the operating procedures.
• Personnel performing experiments who have undergone professional training must wear protective equipment and conduct their operations within a laboratory equipped with appropriate safety safeguards.
• The reagent kit should be stored away from light to prevent degradation of the fluorescent compounds. All consumables used, including centrifuge tubes and tips, must be autoclaved and free of DNase and RNase.
• The test samples involved in this kit shall be regarded as containing infectious substances; all operations and handling procedures must comply with the relevant requirements of the "General Biosafety Guidelines for Microbial and Biomedical Laboratories" issued by the Ministry of Health and the "Regulations on the Management of Medical Waste."