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The monkeypox outbreak has become increasingly severe, and molecular diagnostic techniques are crucial for the early and rapid diagnosis of the monkeypox virus. Key methods include real-time fluorescent quantitative technology, NGS sequencing, and loop-mediated isothermal amplification (LAMP).
Real-time Fluorescent Quantitative Technology
Real-time polymerase chain reaction (RT-PCR) utilizes specific fluorescent-labeled probes for real-time detection of amplification products, providing both high sensitivity and specificity. When genomic fragments of the monkeypox virus are detected in skin lesion samples from infected patients, this confirms a positive diagnosis, establishing RT-PCR as the gold standard for MPXV confirmation. RT-PCR allows for accurate quantitative analysis of multiple samples and has been widely applied in the field of pathogen molecular diagnosis.
NGS Sequencing Technology
High-throughput sequencing (NGS) can directly obtain the nucleic acid sequence of MPXV from monkeypox samples, representing a novel sequencing method that does not rely on traditional culture results.
NGS technology has the advantages of high throughput, low detection limit, high accuracy and rich information. NGS aids in better understanding the epidemiology, sources of infection, and transmission routes of MPXV by analyzing its DNA sequences.
Given its sensitivity and detection speed, NGS is expected to become a promising technology for precise diagnosis and tailored treatment in clinical infectious diseases.
Loop-mediated Isothermal Amplification (LAMP)
LAMP is a new, isothermal nucleic acid amplification technology that does not require thermal cycling, characterized by its efficiency, simplicity, high specificity, and low cost compared to traditional RT-PCR. LAMP detection of MPXV involves designing four primers targeting six specific regions of the viral genome.
Using BstDNA polymerase, which has strand-displacement activity, LAMP catalyzes the synthesis of new strands at constant temperature, resulting in efficient target gene amplification.
Ongoing advancements in LAMP technology are crucial for boosting true positive rates. By integrating this technology with various detection methods, we can further enhance accuracy, making it more suitable for inspections and quarantine procedures at ports of entry and primary healthcare facilities.
Synbio Technologies | Monkeypox Virus Primers and Probes
Compared with biochemical diagnostics and immunodiagnostics, molecular diagnostics provide advantages of shorter detection time, higher sensitivity, and superior specificity. Synbio Technologies’ Syno C synthesis platform has launched several products for monkeypox virus primers and probes, achieving a purity of up to 95% and are available as standard offerings for quick delivery.
Synbio Technologies supports the development and production of molecular diagnostics reagents, providing high-quality raw material solutions for rapid monkeypox virus detection.
| Panel | Cat. No. | Name | Sequence | nmol/Tube | Price |
|---|---|---|---|---|---|
| Generic Monkeypox Virus | HXMPXV-F-20 | MPXV-F（Monkeypox Virus F Primer） | 5’-GGAAAATGTAAAGACAACGAATACAG-3’ | 20 | $119/set |
| HXMPXV-R-20 | MPXV-R（Monkeypox Virus R Primer） | 5’-GCTATCACATAATCTGGAAGCGTA-3’ | 20 | ||
| HXMPXV-P-5 | MPXV-P（Monkeypox Virus R Probe） | 5’-FAM-AAGCCGTAATCTA(T-BHQ1)GTTGTCTATCGTGTCC-SpacerC6-3’ | 5 | ||
| Human DNA (e.g., RNase P) | HXRNP-F-20 | RNP-F（Human RNase P F Primer） | 5’-AGATTTGGACCTGCGAGCG-3’ | 20 | $89/set |
| HXRNP-R-20 | RNP-R（Human RNase P R Primer） | 5’-GAGCGGCTGTCTCCACAAGT-3’ | 20 | ||
| HXRNP-P-5 | RNP-P（Human RNase P Probe） | 5’-FAM-TTCTGACCTGAAGGCTCTGCGCG-BHQ1-3’ | 5 | ||
| Orthopoxvirus (non-variola) | HXNVAR-OPXV-F-20 | NVAR-OPXV-F（Orthopoxvirus F Primer） | 5’-TCA ACT GAA AAG GCC ATC TAT GA-3’ | 20 | $119/set |
| HXNVAR-OPXV-R-20 | NVAR-OPXV-R（Orthopoxvirus R Primer） | 5’-GAG TAT AGA GCA CTA TTT CTA AAT CCC A-3’ | 20 | ||
| HXNVAR-OPXV-P-5 | NVAR-OPXV-P（Orthopoxvirus P Probe） | 5’-FAM-CCA TGC AAT A (T–BHQ1) A CGT ACA AGA TAG TAG CCA AC-Phos-3’ | 5 |
Monkeypox Virus Primers and Probes
References
Miao, Qing, et al. "Microbiological diagnostic performance of metagenomic next-generation sequencing when applied to clinical practice." Clinical Infectious Diseases 67.suppl_2 (2018): S231-S240.
