MGI Tech has released a new sequencing package targeting all four dengue virus serotypes, which the company says will greatly improve the efficiency of dengue fever surveillance and control. Through a streamlined workflow, encompassing nucleic acid extraction, library preparation, DNBSEQ sequencing, and data analysis, the solution will help inform public health initiatives and improve health outcomes by facilitating rapid, accurate, and cost-effective detection and typing of dengue viruses, according to the company.
The solution, called the ATOPlex DENV1-4 Targeted Sequencing Package, is designed to support efficient and user-friendly operation, delivering results in as few as 12 hours to aid rapid dengue fever prevention and control.
The release came as the US Centers for Disease Control and Prevention recorded over 12 million reported cases of dengue in the Americas this year, close to triple last year’s total. With no specific antiviral treatment, public health efforts against dengue fever have been primarily focused on vector control and supportive care and guided by monitoring and surveillance.
Hurdles to Dengue Fever Management
Dengue fever, a debilitating mosquito-borne viral illness, poses a significant threat to global health, particularly in tropical and subtropical regions. With an estimated 390 million cases annually and increasing prevalence in urban and semi-urban areas, the disease places a heavy burden on health care systems and economies worldwide. In addition, its complex clinical presentation, ranging from mild fever to life-threatening hemorrhagic fever, makes accurate and timely diagnosis crucial.
Traditional diagnostic methods, such as serological tests and PCR, often have low throughput and can’t detect variation for effective surveillance and outbreak control. However, the rise of high-throughput sequencing has opened new avenues in infectious disease research. By presenting a detailed genetic blueprint of the virus, sequencing can offer unprecedented insights into pathogen evolution, transmission dynamics, and drug resistance in dengue virus.
MGI DENV1-4 solution
The ATOPlex DENV1-4 Targeted Sequencing Package offers a streamlined workflow encompassing nucleic acid extraction, library preparation, DNBSEQ sequencing, and data analysis, according to MGI.
Its library preparation set, which leverages MGI’s proprietary ATOPlex multiplex PCR technology, ensures high sensitivity and specificity, even with low viral loads. It offers flexible sequencing solutions with SE100 for faster turnaround time and PE100 for cost-effectiveness, catering to diverse user needs.
Furthermore, the integrated ATOPlex MultiPCR Microbiome Research Software (ATOPlex MMRS) is designed to simplify data interpretation. Users can obtain accurate typing results based on built-in reference genome sequences and primer information of several different target microorganisms, including dengue virus, coronavirus, mpox virus, respiratory syncytial virus, and any custom input species.
MGI says the solution is easy to operate, incorporating reverse transcription, multiple amplification, and Fast PCR-Free technology, and library preparation can be completed in one tube with simple operation. It is also suitable for detecting low viral loads (>100 copies/RXN), offering rapid software analysis, accurate typing of DENV1-4, and high accuracy in mutation detection.
Performance:
Performance validation by MGI showed a 100 percent success rate in library preparation by MGISP-100, while the output exceeded the quality control standard of 20ng in all cases with an RNA input of 200 ng. Moreover, library preparation using the MGISP-100RS saw enhanced throughput and reduced hands-on time, demonstrating higher library yields compared to manual methods. All resulting DNB concentrations also measured ≥ 8 ng/μL, meeting sequencing quality thresholds.
At the sequencing stage, different DNBs were loaded onto a lineup of DNBSEQ platforms, including the DNBSEQ-G99RS/ARS, DNBSEQ-E25RS and DNBSEQ-G50. The total reads across all platforms met the requirements of more than 80M, 25M, and 100M, respectively. The Q30 of each flow cell was also greater than 95 percent. At the same time, the average quality control values at 1x, 10x, and 100x sequencing coverage on the three platforms all surpassed 99 percent, and the sequencing results produced by the MGISP-100 were comparable to those obtained manually.
Lastly, further clade identification analysis showed that the typing results of dengue fever samples were consistent with known results, validating the package's accuracy and reliability.
Enhancing the Fight Against Dengue Fever
The impact of the ATOPlex DENV1-4 Targeted Sequencing Package extends from the local to the global level, said MGI. In local communities, it can empower healthcare providers with the tools to rapidly diagnose and manage dengue fever cases, minimizing severe complications and mortality, according to MGI Tech.
Since late 2023, the solution has been deployed in a pilot program in the Chinese cities of Shantou, Jieyang, and Chaozhou, demonstrating its applicability in real-world monitoring efforts. In fact, sequencing results revealed the presence of different dengue virus serotypes circulating in the eastern Guangdong region, highlighting the importance of accurate typing for effective outbreak control.
At the global level, the package is said to contribute to enhanced surveillance networks, enabling real-time monitoring of dengue virus evolution and transmission patterns.