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Monkeypox virus is a kind of enveloped double stranded DNA virus, which is rectangular in shape and can be cultured and grown in African green monkey kidney cells, causing cytopathic effects. It belongs to the orthopoxviral genus of the poxviridae family. Monkeypox is a zoonotic infectious disease caused by monkeypox virus primers. Its clinical manifestations include high fever, headache, general discomfort, cough, enlarged lymph nodes, accompanied by a rash similar to smallpox throughout the body, and occasional abdominal pain. This kit uses fluorescence PCR method and can be applied to Bioer’s LineGene 9600 Plus (FQD-96A), Bioer’s QuantGene 9600 (FQD-96C) and ABI 7500. It has the characteristics of fast detection speed, good specificity, and high sensitivity.

This kit selects two highly conserved regions of monkeypox virus, the F3L gene (FAM) and the B6R gene (HEX) and designs two pairs of specific primer probes. The primers and probes can specifically bind to the target sequence. Under the action of Taq enzyme, the fragment is specifically amplified, and the fluorescent probe is hydrolyzed during the amplification process to produce fluorescence. The detection system contains dUTP-UDG enzyme anti-pollution measures to fully degrade possible product contamination and avoid false positive results. In addition, internal standards are set up to monitor the entire process of sample collection, transportation, nucleic acid extraction, and PCR amplification, avoiding false negative results, and ensuring the effectiveness of the entire process.

Product features
Product parameters
Experimental data

1. Accuracy: After dissolving the positive reference P1-P8 and negative reference N1-N8 of Hangzhou Bioer Technology Co., Ltd. as required, MagaBio plus Virus DNA/RNA Purification Kit III BSC86S1E (ZHXB No. 20200872) was used to extract the nucleic acid.

Figure 1: Detection of negative/positive reference materials in monkeypox virus reference materials

The results showed that the nucleic acid of monkeypox virus could be accurately detected. Test results of enterprise reference materials: positive coincidence rate 100%, negative coincidence rate 100%.

2. Precision: After re-dissolving the precision reference J1-J2 of the Monkeypox Virus Nucleic Acid Multiple Detection Kit (Fluorescent PCR) according to the usage requirements, extract it with MagaBio plus Virus DNA/RNA Purification Kit III BSC86S1E (ZHXB No. 20200872), use three batches of reagents for testing, and repeat the test for 10 times.

Figure 2  Intra-assay and inter-assay precision

The results showed that the intra-assay and inter-assay precision variation coefficients of the three batches of reagents were less than 5%, and the reagent precision was good.

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