Domestic gene sequencer reproduces new breakthroughs.

Recently, a reporter from 21st century business herald learned that the fourth-generation gene sequencing technology "unicorn" Anxenyuan has released the fourth-generation high-throughput nanopore gene sequencer (AXP100-RS) to the public. Its startup cost is as low as 100 yuan, which has opened the prelude to the development of low-cost long-term reading in the sequencing industry.

tu haobo, director of anxuyuan products, pointed out that the key to the ultra-low cost of the product lies in the company's unique large throughput chip based on CMOS process while maintaining long reading, high throughput and high accuracy. "we have now introduced the world's only 12-inch nanopore sequencing wafer in mass production. this chip has entered the stage of stable mass production, with a single chip producing data of 100Gb, the cost of a single chip is much lower than US $100, and the sequencing cost is as low as US $0.3/Gb."

it is worth mentioning that among the current mainstream manufacturers of the second generation gene sequencers, the price of each chip in Illumina is more than us $10,000, and that of huada zhizao (688114.SH) is also in thousands of us dollars.

In the view of some market participants, with the further maturity of the fourth-generation gene sequencer technology, it is expected to make up for the shortcomings of the second-generation sequencer such as long reading, complex processes, and large instrument size, expand the application scale of sequencing technology, And rely on lower costs to further sink to the grassroots market.

Image source: Visual China

Gene sequencing technology "four generations together"

Gene is the underlying code of the "central law" of life sciences. In recent years, the demand for the middle and downstream of the gene sequencing industry chain has continued to grow, and the upstream gene sequencer has become one of the hottest tools in the field of life sciences.

At present, the mainstream gene sequencing technology "four generations in the same house" exists in parallel. Each sequencing technology has its own strengths, but each has its own shortcomings. From the scientific research breakthroughs and downstream application needs in recent years, long-read long sequencing is an industry trend, limited by the impact of high costs, long-read long technology applications still account for a small proportion.

The first generation of sequencing is the Sanger method, which is characterized by long read length and high accuracy, but low throughput and high cost, which makes it difficult to meet the data volume requirements of modern credit analysis models.

Second-generation sequencing (NGS), the dominant Illumina of global sequencers, quickly enters the market with high-throughput and high-accuracy technical characteristics, and promotes the rapid decline of gene sequencing costs, and occupies a monopoly position with relative cost advantages For more than ten years, it has played a pivotal role in the process of gene sequencing penetrating from high-precision scientific research services into clinical applications.

At present, the global market is generally dominated by second-generation gene sequencing technology (NGS), accounting for nearly 80% of the market share. The domestic enterprise Huada Zhizao (688114.SH) acquired the core patented technology of sequencers through the acquisition of CG company, and also broke through the domestic problem of sequencers.

However, the second-generation gene sequencer is not a panacea. Its reading length, complicated process and difficult operation cannot cover the sequencing needs of complex samples in a wider range of clinical scenarios.

"Although the cost of the second generation sequencing technology is significantly lower than that of the first generation, the cost is still very high for large-scale application in the clinical end. First of all, sequencing instruments generally cost 400000-700000 US dollars, and a whole genome sequencing requires at least 2000-5000 US dollars. Then there is the time cost. The second generation sequencing technology relies on the amplification process of gene samples. A large number of elution processes not only increase the cost and sample preparation time, but also are prone to error accumulation. The third is the read length. The second generation sequencing technology generally has a read length of 150-400bp (base pairs), which cannot meet the needs of higher scientific research." A partner of a third-party service organization in the field of life sciences in South China pointed out to this reporter.

Therefore, in recent years, the three-generation and four-generation sequencing technologies based on the principle of single-molecule fluorescence sequencing and single-molecule nanopore sequencing have received widespread attention in the market in recent years due to the characteristics of ultra-long reading length, flexibility and convenience.

Pacific Biosciences (hereinafter referred to as "Pacbio"), the leading enterprise of the third generation sequencing technology, has been listed on NASDAQ in the United States. It is a single molecule real-time sequencing method based on zero mode waveguide, with high accuracy. However, the sequencing price is high and the application scope is limited.

The fourth generation sequencing technology is called nanopore sequencing, which is a technology based on electrical signal sequencing, which can identify the base information on each nucleic acid molecule according to the current signal, so as to realize the sequencing of single-stranded nucleic acid molecules.

"the main advantage of the fourth generation sequencer is that it can achieve ultra-high reading length, which can reach 10kb to hundreds of kb (kilobase pairs), and does not require elution and PCR amplification, direct sequencing and real-time acquisition of sequence information, with relatively low equipment cost." The aforementioned third-party service partner further pointed out.

in overseas markets, Roche acquired Genia, a fourth-generation sequencing technology company, as early as 2014. Illumina also focused on long-reading and long-term sequencing technology. in order to make up for its NGS short board, Roche invested in Oxford Nanopore (Oxford nanopore company, hereinafter referred to as "ONT"), a global nanopore sequencing leader. In September 2021, ONT was successfully listed on the London Stock Exchange, and its market value soared to $6.8 billion on its first day. However, the company is not yet profitable and is expected to break even in 2026.

In the domestic market, all kinds of capital have also laid out the fourth generation sequencing technology.

in March 2023, a-share listed company wanfu biology (300482.SZ) announced that it plans to invest 30 million yuan with its own funds to invest in Beijing puyi biotechnology co., ltd. the latter focuses on the development of a new generation of nucleic acid and other biopolymer sequencers and supporting reagent consumables based on biological nanopores.

Wanfu Biology pointed out in the announcement: "Nanopore sequencing technology is expected to bring new breakthroughs in clinical application scenarios such as tumor accompanying diagnosis and early screening, genetic disease and rare disease diagnosis, cell and gene therapy, etc. in terms of reading length, speed and epigenetics."

"At present, various nanopore sequencing technologies have solved the problem of long read length, and there is still a breakthrough in terms of accuracy and cost. Throughout the history of the industry, the ultimate trend of sequencing technology will be low-cost long read length." A senior fourth-generation sequencing technology people told reporters.

An sequence source breaks the situation

Although the development of fourth-generation gene sequencing technology is still early, some head players have also emerged in the market.

among them, the relatively mature single-molecule nanopore sequencer comes from ONT. in 2014, the first commercial device MinION (portable sequencer) of ONT was born, which has attracted market attention due to its advantages such as "u disk type", "ultra-long reading length", "real-time results" and "on-demand sequencing. Since then, ONT has continuously expanded its product line and introduced sequencing equipment of different sizes such as GridION, GridION X5 desktop sequencer and high-throughput sequencer PromethION.

however, huada zhizao prospectus believes that "ONT's fourth generation nano sequencer has a longer read length than high-throughput sequencing technology, but its single base error rate is about 1%-10%, which is higher than that of high-throughput sequencing technology."

when ONT went on the market in October 2021, the original reading accuracy of DNA/RNA data of its sequencing equipment has increased from 80% in 2014 to 99.8, but the market generally believes that the accuracy of the fourth generation ONT sequencer still has room for further improvement.

Tu Haobo, director of anserom source products, told reporters that as a representative company of the fourth generation gene sequencer in China, the technical path of anserom source is CCS + nanopore single molecule sequencing, which combines the advantages of the third generation Pacbio sequencer and the fourth generation ONT sequencer. The miniaturized mobile single molecule sequencing platform based on nanopore chips can have the characteristics of high throughput, long reading length, high precision and low cost.

"the fourth generation sequencer AXP 100-RS currently produces good data quality. the core competitive advantages of the source are mainly three points: first, the cost and throughput advantages brought by high-density chips, I .e. lower cost and greater throughput; Second, the accuracy can be significantly improved by adopting cyclized sequencing technology. In addition, we use the form of alternating current, multi-dimensional measurement, clear signal, is the real single molecule, single base sequencing." Tu Haobo further said.

The new AXP100-RS sequencer released this time will be used in genetic diseases, infectious diseases, cancer and other fields in the future. Tian Hui, CEO of Anxuyuan, pointed out at this product conference: "Sequencing technology has been fruitful in the past few decades., The market is huge. However, due to the constraints of start-up cost and data output cost, the field of sequencing application is still very limited. Even in the key application areas that have been developed, such as tumor detection, sequencing can still be said to be in the embryonic stage."

judging from the performance of sequencing upstream manufacturers and the application product layout of sequencing downstream enterprises, the price decline of the whole sequencing industry has entered a bottleneck period, and new revolutionary sequencing products are needed to lead the industry direction.

our reporter learned from the scene of anxuyuan's press conference that the company's greatest technical advantage is biochips, which are based on mass production of large wafers and can reduce the sequencing startup cost to 100 yuan, and the unit data output cost is far lower than NGS. in the future, the sequencing cost can be further reduced through technological upgrading.

With the decline in the price of sequencing, it will bring a great expansion of downstream applications. According to relevant data released by illumina at the 41st JP Morgan JPMorgan Medical and Health Industry Annual Meeting, it is estimated that the total target market for global sequencing applications will reach US $120 billion in 2027.

"We will start with gene sequencing and inject more hard technology into the field of in vitro diagnostics to create an IC + BT underlying platform for the big health industry. Through the price advantage of the mass production process, we will bring the sequencing of everything at our fingertips and open up new horizons to recognize the origin of life." Tian Hui said.