NEW YORK – A new sample preparation approach developed by Atila Biosciences could potentially enable affordable, easy-to-use, and high-throughput global infectious disease testing.

纽约——Atila Biosciences开发的一种新的样品制备方法可能会使负担得起，易于使用和高通量的全球传染病检测成为可能。

Atila's Biodome technology is a novel hydrophobic gel matrix that sequesters liquid PCR master mix, primers, and enzymes within a single reaction tube so that they don't mix during shipping yet melts during thermocycling and floats to the surface of the reaction.

Atila的Biodome技术是一种新型疏水凝胶基质，可将液体PCR主混合物，引物和酶隔离在单个反应管中，以便它们在运输过程中不会混合，但在热循环过程中会融化并漂浮到反应表面。

The tech has been adopted by a team based in the Feinberg School of Medicine at Northwestern University and is now helping global health researchers broaden their screening and cancer prevention programs in low- and middle-income countries while also reducing expert staffing and downtime caused by contamination.   .

这项技术已被西北大学范伯格医学院的一个团队采用，目前正在帮助全球卫生研究人员在中低收入国家扩大筛查和癌症预防计划，同时减少专家人员配备和污染造成的停工时间。。

'It really is a game changer,' said Mamoudou Maiga, director of translational research for Northwestern's Center for Innovation in Global Health Technologies (CIGHT), who has been using Biodome for human papillomavirus screening in Mali and Nigeria. 'It's very a simple thing that solves big issues,' he said..

西北大学全球卫生技术创新中心（CIGHT）转化研究主任马穆杜·麦加（MamoudouMaiga）说，这确实是一个游戏规则的改变者，他一直在马里和尼日利亚使用Biodome进行人乳头瘤病毒筛查。他说，这是一件非常简单的事情，可以解决大问题。。

Contamination is a problem for any clinical microbiology lab since each PCR reaction generates millions of amplicons that can potentially interfere with subsequent runs of extremely sensitive molecular tests.

污染对于任何临床微生物学实验室来说都是一个问题，因为每个PCR反应都会产生数百万个扩增子，这些扩增子可能会干扰随后进行的极其敏感的分子检测。

'It makes the space very dirty quickly, and there is no way to avoid that,' Maiga explained. Labs typically have separate clean lab and dirty lab spaces to try to prevent contamination, but contamination looms at each step in the pre-amplification workflow. Lyophilizing nucleic acid amplification reagents, meanwhile, can help increase their shelf life and allow for room-temperature shipping but requires clean, nuclease-free water for rehydration..

麦加解释说：“这很快会让空间变得非常脏，这是无法避免的。”。实验室通常有单独的干净实验室和脏实验室空间，以防止污染，但在预放大工作流程的每个步骤都会出现污染。同时，冻干核酸扩增试剂可以帮助延长其保质期并允许室温运输，但需要干净的无核酸酶水进行再水化。。

Biodome hermetically seals reagents, however, so they can be shipped at room temperature while technicians need only pipette their sample on top prior to testing.

然而，Biodome密封试剂，因此可以在室温下运输，而技术人员只需在测试前将样品移液到顶部即可。

The semi-solid gel matrix melts above 45 degrees Celsius, and because it is hydrophobic it then floats to the surface to mix the reagents as thermal cycling begins. It also solidifies on the surface at the end of PCR, re-sealing the reaction and preventing post-amplification contamination.

半固体凝胶基质在45摄氏度以上熔化，由于它具有疏水性，因此在热循环开始时会漂浮到表面以混合试剂。它还在PCR结束时在表面固化，重新密封反应并防止扩增后污染。

Lifang Hou, the director of the Center for Global Oncology at Northwestern's Institute for Global Health, said troubleshooting lab contamination is costly and time consuming, noting that in the past her team had to shut down a project in Nigeria for several months to find a contamination source, a process itself that also consumed valuable reagents.

西北大学全球卫生研究所全球肿瘤学中心主任侯丽芳表示，排除实验室污染既费钱又费时，她指出，过去她的团队不得不关闭尼日利亚的一个项目数月，以寻找污染源，这一过程本身也消耗了宝贵的试剂。

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'We had to stop recruitment,' she said, which in her line of work is particularly painful since she is trying to catch infections to stop the spread of diseases that can cause fatal cancers.

“我们不得不停止招聘，”她说，这在她的工作领域尤其痛苦，因为她正试图感染以阻止可能导致致命癌症的疾病传播。

Maiga, Hou and their global health colleagues used assays from Cepheid in research

Maiga，Hou和他们的全球健康同事在研究中使用了造父变星的检测方法

published

已发布

last year, showing that for a sample of 96 women living with HIV, 42 percent were also infected with high-risk HPV, a prevalence generally consistent across sub-Saharan Africa. It is now known that almost 99 percent of cervical cancer cases are linked to HPV infection, Maiga said, and, 'Without HPV, there is no cervical cancer.'.

去年的研究表明，在96名感染艾滋病毒的女性样本中，42%的人也感染了高危型HPV，这种流行率在撒哈拉以南非洲地区基本一致。麦加说，现在已经知道，几乎99%的宫颈癌病例与HPV感染有关，“没有HPV，就没有宫颈癌。”。

Unlike lower-throughput methods that prevent contamination through the use of cartridges, the Atila HPV testing approach enables high-throughput testing at a lower cost.

与通过使用墨盒防止污染的低通量方法不同，Atila HPV检测方法能够以较低的成本进行高通量检测。

The Biodome tech is available in tube strips or a plate-based format and can be used with Atila's AmpFire and ScreenFire diagnostics systems though it is essentially instrument-agnostic, Maiga and Hou said, and they have tested it with four other thermal cyclers.

Maiga和Hou说，Biodome技术可以以管带或平板形式提供，可以与Atila的AmpFire和ScreenFire诊断系统一起使用，尽管它基本上与仪器无关，他们已经用其他四个热循环仪进行了测试。

HPV screening and beyond

HPV筛查及其他

Awarded nearly $2 million to date by the National Cancer Institute for studies of HPV screening and self-sampling in Western Africa, Maiga and Hou have found that allowing people to collect their own vaginal swabs overcomes many cultural and religious barriers to gynecological exams.

Maiga和Hou发现，允许人们收集自己的阴道拭子克服了妇科检查的许多文化和宗教障碍，迄今为止，该研究所获得了西非国家癌症研究所颁发的近200万美元的奖金。

A single technician can collect swabs from 100 women in five minutes, Maiga noted, and the 96-well Atila test format facilitates more than 1,000 tests per day. This allows technicians to triage the 20 percent or so of women who have high-risk HPV for additional care while sending the rest home.

Maiga指出，一名技术人员可以在五分钟内从100名女性身上采集拭子，而96孔Atila测试格式每天可以进行1000多次测试。这使得技术人员可以对20%左右的高危型HPV女性进行分流，以便在将其余患者送回家的同时进行额外护理。

High-risk HPV-positive women require visual inspection of the cervix (VIA), Hou said, which demands a clinician time and a private exam room. 'You can imagine if you use Atila's testing to understand who is in need of VIA, how much public and clinical resources will be saved,' she said.

侯说，高危型HPV阳性女性需要目视检查宫颈（VIA），这需要临床医生的时间和私人检查室她说：“你可以想象，如果你使用Atila的测试来了解谁需要VIA，将节省多少公共和临床资源。”。

Lives will also be saved, she suggested, and the fact that HPV is now treatable makes ongoing high cervical cancer deaths rates globally an issue of health equity and justice. 'That is why we are so devoted to this work,' Hou said.

她建议说，生命也将得到挽救，而HPV现在可以治疗的事实使得全球宫颈癌死亡率居高不下，成为健康公平和正义的问题侯说，这就是为什么我们如此专注于这项工作。

The team plans to port its HPV screening efforts to mobile clinics using two Atila systems and the Biodome technology. The method isn't just for HPV, Hou said, adding her team is using it for malaria testing in Mali and will soon use it for tuberculosis testing.

。Hou说，这种方法不仅适用于HPV，她的团队正在马里使用它进行疟疾检测，不久将用于结核病检测。

The team runs a training program for clinicians in Mali and Nigeria, as well as a network of HPV cervical cancer programs in seven countries in Africa, including Burkina Faso, Senegal, Guinea, Nigeria, Tanzania, Rwanda, and Kenya, Maiga said. In these programs, too, Maiga and his colleagues are expanding their portfolio to diseases beyond HPV using the Atila system.

麦加说，该团队在马里和尼日利亚为临床医生举办了一个培训项目，并在非洲七个国家（包括布基纳法索、塞内加尔、几内亚、尼日利亚、坦桑尼亚、卢旺达和肯尼亚）建立了HPV宫颈癌项目网络。在这些项目中，Maiga和他的同事也在使用Atila系统将他们的投资组合扩展到HPV以外的疾病。

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For populations in the regions, 'Want to do affordable, scalable molecular testing in their own country, not sending tests to Europe,' Hou said.

侯说，对于这些地区的人群来说，他们希望在自己的国家进行负担得起、可扩展的分子检测，而不是将检测发送到欧洲。

Maiga and Hou recently validated the Biodome tech on samples from Mali and Nigeria processed at their labs in Northwestern, with results reported in an

Maiga和Hou最近在西北部实验室处理的马里和尼日利亚样品上验证了Biodome技术，结果报告在

Infectious Agents and Cancer

传染源与癌症

study published late last year. Specifically, the team compared the Biodome format in Atila's isothermal ScreenFire HPV RS assay to the results of using the ScreenFire HPV RS assay standard format on Biorad CFX-96 real-time PCR instrument.

这项研究于去年年底发表。具体来说，该团队将Atila等温ScreenFire HPV RS检测中的Biodome格式与在Biorad CFX-96实时PCR仪器上使用ScreenFire HPV RS检测标准格式的结果进行了比较。

ScreenFire HPV RS employs Atila's proprietary isothermal amplification and 96-well format to detect high-risk HPV with a sensitivity of approximately 95 percent, as

ScreenFire HPV RS采用Atila专有的等温扩增和96孔格式检测高危HPV，灵敏度约为95%

previously reported

以前报告过

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Specifically, the test uses four fluorescent detection channels with sensitivity hierarchically tailored to cancer risk, such that the most sensitive component assays detect HPV16 and HPV18/45, while less sensitive assays detect HPV strains 31/33/35/52/58, and HPV strains 39/51/56/59/68.

具体而言，该测试使用四种荧光检测通道，其灵敏度分层定制为癌症风险，因此最敏感的成分测定法检测HPV16和HPV18/45，而较不敏感的测定法检测HPV株31/33/35/52/58和HPV株39/51/56/59/68。

However, without the Biodome sample prep tech, ScreenFire HPV RS still requires manual preparation of master mix reagents, the team wrote in the study.

然而，在没有Biodome样品制备技术的情况下，ScreenFire HPV RS仍然需要手动制备预混试剂，该团队在研究中写道。

For the HPV RS test, Atila developed what it calls Zebra BioDome — a preloaded format that stacks and separates the reagents needed for its proprietary tests.

对于HPV RS测试，Atila开发了一种称为Zebra BioDome的预加载格式，可以堆叠和分离其专有测试所需的试剂。

In the study, the Northwestern team set up a Bio-Rad Laboratories CFX-96 real-time PCR machine with a test program of one minute per cycle at 60

在这项研究中，西北大学的研究小组建立了一台Bio-Rad实验室CFX-96实时PCR仪，测试程序为60

C

C级

for 60 cycles. They found the HPV screening workflow using Zebra Biodome had approximately 98 percent agreement with the standard workflow but reduced the number of pipetting steps from three to one.

进行60个循环。他们发现使用Zebra Biodome的HPV筛查工作流程与标准工作流程的一致性约为98%，但将移液步骤的数量从三个减少到一个。

In an assessment of 173 banked samples of extracted nucleic acids, only seven samples showed inconsistent HPV genotyping results between the two tests. These were later found to have viral loads near the limit of detection, and increasing the sample volume from five microliters to 10 microliters led to complete agreement between the two tests.

在对173个提取的核酸样本库的评估中，只有7个样本在两次测试之间显示出不一致的HPV基因分型结果。后来发现这些病毒载量接近检测限，将样本量从5微升增加到10微升导致两次测试完全一致。

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In addition, the Northwestern team validated the Biodome tech in 46 crude samples that had been collected previously. They added vaginal swabs to lysis buffer before pipetting into the assays, finding 100 percent agreement between Biodome and the standard format.

此外，西北大学团队在之前收集的46个原油样本中验证了Biodome技术。他们将阴道拭子加入裂解缓冲液中，然后移液到测定中，发现Biodome与标准格式之间100%一致。

Other work published in the

其他发表在

Journal of Clinical Microbiology

临床微生物学杂志

in December by researchers based at the National Cancer Institute and in Nigeria showed similar agreement between Biodome and the standard Atila formats. This validation study built upon prior work comparing Atila's HPV testing approach to the Cepheid GeneXpert as well, showing a $6 per test compared to $15 per test cost, and noting that the technology can have broad applications for other DNA amplification assays beyond HPV..

去年12月，国家癌症研究所和尼日利亚的研究人员在Biodome和标准Atila格式之间达成了类似的协议。这项验证研究建立在先前将Atila的HPV检测方法与Cepheid GeneXpert进行比较的基础上，显示每次检测6美元，而每次检测成本为15美元，并指出该技术可以广泛应用于HPV以外的其他DNA扩增检测。。

Atia's plans

Atia的计划

Youxiang Wang, CEO and founder of Atila, said in an interview that the company spent about two years developing Biodome and now hopes to sell preloaded Zebra Biodome assays going forward as well as offer a service for assay developers to port their tests to the universal Biodome format.

Atila首席执行官兼创始人王友祥（Youxiang Wang）在接受采访时表示，该公司花了大约两年的时间开发Biodome，现在希望销售预装的斑马Biodome检测方法，并为检测开发者提供服务，将其检测结果移植到通用Biodome格式。

'My vision is that this format will replace lyophilization,' Wang said, a process that he noted is tedious and impacts production capacities and timelines. The addition of Biodome matrix to tubes or plates is automated, he said, and only adds a few cents to the cost of an assay.

“我的愿景是，这种形式将取代冻干，”王说，他指出，这个过程很乏味，会影响生产能力和时间表。他说，将Biodome基质添加到试管或平板中是自动化的，只会增加检测成本几美分。

Atila also hopes to partner with larger companies to make the Biodome technology more widely available in their consumables, he said, so 'Then they never need to worry about lab contamination.'

他说，Atila还希望与较大的公司合作，使Biodome技术在他们的消耗品中更广泛地使用，这样“他们就不必担心实验室污染”

Atila was recently granted breakthrough device designation by the US Food and Drug Administration for use of the ScreenFire HPV RS assay in anal specimens for anal HSIL and anal cancer screening of HIV-positive individuals over 35 years of age.

Atila最近被美国食品和药物管理局授予突破性的设备指定，用于在肛门标本中使用ScreenFire HPV RS检测，用于对35岁以上HIV阳性个体进行肛门HSIL和肛门癌筛查。

The tech may also help meet global goals to reduce cancer rates in LMICs. With a low throughput, 'Point-of-care testing is never going to finish screening 70 percent of the women in the world,' Wang said, adding that cartridge-based tests prevent contamination but are typically more costly than lab-based testing.

这项技术也可能有助于实现降低中低收入国家癌症发病率的全球目标。Wang说，由于吞吐量低，“即时检测永远无法完成对世界上70%的女性的筛查”，并补充说，基于墨盒的检测可以防止污染，但通常比基于实验室的检测成本更高。

Yet, throughput using Atila's approach is also modifiable, he said, as users can trim down tube strips to run just two samples if needed. .

然而，使用Atila方法的吞吐量也是可以修改的，他说，因为用户可以根据需要修剪管带，只运行两个样本。

'This innovation is going to change the field,' Wang said.

王说，这项创新将改变这个领域。