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The development of COPMAN-Air: A highly sensitive method for detecting SARS-CoV-2 in air
Thursday February 20th, 2025 at 1:59 PM

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Page 1/13
The development of COPMAN-Air: A highly sensitive
method for detecting SARS-CoV-2 in air
Tomoyo Yoshinaga
Shionogi & Co., Ltd
Yoshinori Ando
Shionogi & Co., Ltd
Yumi Sato
Shionogi & Co., Ltd
Takeru Kishida
Kishida Clinic
Masaaki Kitajima
The University of Tokyo
Article
Keywords: Air sampling, COPMAN, COPMAN-Air, Fever clinic, qPCR, SARS-CoV-2
Posted Date: February 18th, 2025
DOI: <a href="https://doi.org/10.21203/rs.3.rs-5995479/v1" rel="nofollow">https://doi.org/10.21203/rs.3.rs-5995479/v1</a>
License: This work is licensed under a Creative Commons Attribution 4.0 International License.
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Additional Declarations: Competing interest reported. Tomoyo Yoshinaga, Yoshinori Ando, and Yumi Sato
are employees of Shionogi & Co., Ltd. Masaaki Kitajima received research funding and patent royalties
from Shionogi & Co., Ltd.

Page 2/13
Abstract
Several studies have successfully detected SARS-CoV-2 in air samples; however, in most of these, the
focus was on validating the air collection method, and there was no report on the development of a
virus-detection method. In this study, to detect viruses in air samples more sensitively than conventional
detection methods, we applied COPMAN, a highly sensitive virus-detection method using wastewater
samples, to air samples to develop COPMAN-Air. Briey, with this method, the extremely low amount of
viral RNA in air samples is eciently detected via three reaction steps: RT, preamplication, and qPCR, as
with COPMAN. We evaluated COPMAN-Air using samples from a fever clinic for COVID-19 patients.
COPMAN-Air demonstrated a higher detection rate of viral RNA compared to conventional methods: 22
(95.7%) vs. 14 (60.9%) out of 23 samples. Additionally, a positive correlation (r=0.70) was found between
the amount of viral RNA detected by COPMAN-Air and the number of conrmed COVID-19 cases,
suggesting that COPMAN-Air could estimate the number of SARS-CoV-2-positive individuals in a given
space based on the quantitative values of SARS-CoV-2 RNA in air samples. Surveillance systems for
pathogens in the air using COPMAN-Air are expected to be valuable for assessing the number of
infected individuals and for the implementation of public health measures.
Introduction
SARS-CoV-2 is a virus that infects the human respiratory tract. It can be spread not only through direct
and indirect contact, but also through the inhalation of droplets and aerosols 1–6. Generally, people
infected with COVID-19 exhibit symptoms similar to those of the common cold or inuenza and recover
after several days, except for vulnerable people such as the elderly and those with underlying health
conditions 7,8. In addition, it has been reported that some people infected with SARS-CoV-2 show no
symptoms, and it is possible that these asymptomatic people may inadvertently play a role as spreaders
of the virus 9–12. Because environmental tests, which detect pathogens contained in the environment,
are non-invasive and are regarded as more cost-effective than clinical tests, they are attracting attention
as an alternative or complement to clinical testing 13.
SARS-CoV-2 is widely accepted as a virus that spreads through airborne transmission; therefore, one way
to prevent its spread is to help people to know if the air around them contains the virus. Air sampling to
detect the virus has been vigorously taken into consideration. Many of these studies have been
conducted in healthcare settings, such as isolation rooms for COVID-19 patients in hospitals and
quarantine hotels, where high concentrations of SARS-CoV-2-containing aerosols are expected to ll the
space 6,14–16. However, considering the social implementations of the virus, it is necessary to conduct a
feasibility study in a more practical community setting. Some reports have successfully detected SARS-
CoV-2 in public places such as student dormitories, schools, cafeterias, oces, shopping centers,
airports, and public transport 16–19. Nevertheless, because it is dicult to grasp the number of SARS-
CoV-2-infected individuals who have been or are presently in such spaces during air sampling, it is not

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possible to accurately conrm the validity of the virus detection method. Furthermore, it cannot be
determined whether the method is available for the early detection of infected individuals.
The major detection method for SARS-CoV-2 in the air is the quantitative measurement of viral RNA
derived from an air sample via reverse transcription-quantitative polymerase chain reaction (RT-qPCR)
15,16,18. There have been many reports on air sampling methods 16,20. Since the number of SARS-CoV-2-
containing aerosols in a space is expected to be extremely low, the optimization of the protocol to be
used after air sampling is also needed, to improve sensitivity of virus detection using RT-qPCR. In
addition, the protocol must be able to handle a large number of air samples for virus monitoring in many
spaces, to achieve worthwhile social implementation. To our knowledge, however, such efforts have not
yet been suciently veried.
Recently, our group developed the COPMAN (COagulation and Proteolysis method using MAgnetic
beads for Nucleic acids in wastewater) method, which can detect SARS-CoV-2 RNA from wastewater
samples with both high sensitivity and high throughput 21,22. In this study, we developed COPMAN-Air, a
new method that applies the near full-automation-available COPMAN technology and investigated the
detection sensitivity of SARS-CoV-2 RNA derived from air samples taken from a Thermo Fisher Scientic
AerosolSense Sampler. Some reports have clearly shown that this active air sampler is helpful in
collecting SARS-CoV-2-containing aerosols; however, considering the reported sensitivity and the
throughput estimated from the method used, there could still be room for improvement in the detection
method for social implementation. Furthermore, using our method, we measured the amount of SARS-
CoV-2 RNA in air samples collected at a so-called “fever clinic,” where outpatients with cold symptoms
are examined.
Results
Development of the COPMAN-Air method
We developed a new method (COPMAN-Air) for extracting nucleic acids from the aerosol-absorbed
media collected by the AerosolSense sampler. COPMAN-Air is based on the COPMAN method and
allows for the quantitative measurement of the amount of SARS-CoV-2 RNA in air samples. Additionally,
we conrmed its ability to detect SARS-CoV-2 RNA from a sampler spiked with inactivated SARS-CoV-2
using the COPMAN-Air method.
To compare the sensitivity of COMPAN-Air with the conventional method, we assessed its ability to
detect and quantify SARS-CoV-2 RNA from media spiked with 50, 100, and 1000 copies of inactivated
SARS-CoV-2. The theoretical limit of detection (LOD) of the COPMAN-Air test was lower than
conventional methods, which use 5 µL of RNA, as 14 µL of RNA are subjected to qPCR detection with the
COPMAN-Air method. We compared the observed concentrations between the two methods, since both
were able to detect quantiable amounts of SARS-CoV-2 RNA from the media spiked with 1,000 copies
of the virus. COPMAN-Air showed a higher observed concentration (516.2 copies/sampler) than the

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conventional method (319.5 copies/sampler (N gene), 224.1 copies/sampler (ORF1ab gene), 54.1
copies/sampler (S gene)). A t-test was conducted to compare COPMAN-Air with the conventional
method (N gene) and revealed a signicant difference (
p < 0.05). (Fig. 1a).
The COPMAN-Air method exhibited greater accuracy, compared to the conventional method, because its
coecient of variation was 7.2%, whereas those of the conventional methods were 24.9% (N gene),
21.7% (ORF1ab gene), and 20.3% (S gene).
When spiked with 50 copies of the virus, which is close to the LOD levels, COPMAN-Air exhibited a
greater detection rate than the conventional method (Fig. 1b). COPMAN-Air can be considered a more
sensitive method, compared to the conventional method, due to its lower theoretical LOD and greater
observed concentrations and detection rates.
Validation of COPMAN-Air and its comparison with the conventional method at a fever clinic
To compare COPMAN-Air with the conventional method using eld aerosol samplers, we conducted air
sampling at a fever clinic during the 5th wave of COVID-19 infections in Japan and evaluated these
samples. Based on the measurement results, COPMAN-Air was able to detect SARS-CoV-2 more
accurately in 22 (95.7%) out of 23 samples, with mean concentrations of 1217 copies/sampler, whereas
the conventional method detected the virus in only 14 (60.9%) out of 23 samples (Fig. 2). These ndings
from the clinic experiment led us to conclude that COPMAN-Air demonstrated superior detection
sensitivity than the conventional method. As a result, we have decided to use COPMAN in our future
clinic experiments.
Correlation analysis of SARS-CoV-2 in air samples with the number of COVID-19 patients
COPMAN-Air could detect SARS-CoV-2 even when only a few COVID-19 patients were present (Fig. 2 and
Supplementary Table S1). To compare this with the total number of patients, we conducted extensive
additional air sampling at the fever clinic during the 6th and 7th waves of COVID-19 in Japan
(Supplementary Table S1). We evaluated the correlation between the number of COVID-19 patients and
the amount of SARS-CoV-2 virus RNA detected in the air samples using COPMAN-Air (Fig. 3). The data
points plotted on a scatter plot closely aligned along an approximate straight line (y = 1.066x + 1.590),
suggesting a linear relationship between the number of COVID-19 patients and the viral RNA detected in
the air samples. The results of the Pearson correlation test revealed a positive correlation between the
number of COVID-19 patients and the results of copy numbers of SARS-CoV-2 RNA found the air
samples measured using COPMAN-Air (r = 0.70).
Discussion
In most of the air samples available in this study, we were able to obtain quantitative data for SARS-CoV-
2 RNA using COPMAN-Air. There are two possible reasons why COPMAN-Air showed a higher observed
concentration and lower theoretical LOD compared to the conventional method. Although the protocols

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sarcozona
20 minutes ago
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COOL
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An invisible mask? Wearable air curtain, treated to kill viruses, blocks 99.8% of aerosols
Thursday February 20th, 2025 at 1:43 PM

University of Michigan News
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Headworn tech from U-M startup could protect agricultural and industrial workers from airborne pathogens

An air curtain shooting down from the brim of a hard hat can prevent 99.8% of aerosols from reaching a worker’s face. The technology, created by University of Michigan startup Taza Aya, potentially offers a new protection option for workers in industries where respiratory disease transmission is a concern.

Independent, third-party testing of Taza Aya’s device showed the effectiveness of the air curtain, curved to encircle the face, coming from nozzles at the hat’s brim. But for the air curtain to effectively protect against pathogens in the room, it must first be cleansed of pathogens itself. Previous research by the group of Taza Aya co-founder Herek Clack, U-M associate professor of civil and environmental engineering, showed that their method can remove and kill 99% of airborne viruses in farm and laboratory settings.

“Our air curtain technology is precisely designed to protect wearers from airborne infectious pathogens, using treated air as a barrier in which any pathogens present have been inactivated so that they are no longer able to infect you if you breathe them in,” Clack said. “It’s virtually unheard of—our level of protection against airborne germs, especially when combined with the improved ergonomics it also provides.”

Fire has been used throughout history for sterilization, and while we might not usually think of it this way, it’s what’s known as a thermal plasma. Nonthermal, or cold, plasmas are made of highly energetic, electrically charged molecules and molecular fragments that achieve a similar effect without the heat. Those ions and molecules stabilize quickly, becoming ordinary air before reaching the curtain nozzles.

Taza Aya’s prototype features a backpack, weighing roughly 10 pounds, that houses the nonthermal plasma module, air handler, electronics and the unit’s battery pack. The handler draws air into the module, where it’s treated before flowing to the air curtain’s nozzle array.

Taza Aya’s progress comes in the wake of the COVID-19 pandemic and in the midst of a summer when the U.S. Centers for Disease Control and Prevention have reported four cases of humans testing positive for bird flu. During the pandemic, agriculture suffered disruptions in meat production due to shortages in labor, which had a direct impact on prices, the availability of some products and the extended supply chain.

In recent months, Taza Aya has conducted user experience testing with workers at Michigan Turkey Producers in Wyoming, Michigan, a processing plant that practices the humane handling of birds. The plant is home to hundreds of workers, many of them coming into direct contact with turkeys during their work day.

To date, paper masks have been the main strategy for protecting employees in such large-scale agriculture productions. But on a noisy production line, where many workers speak English as a second language, masks further reduce the ability of workers to communicate by muffling voices and hiding facial clues.

“During COVID, it was a problem for many plants—the masks were needed, but they prevented good communication with our associates,” said Tina Conklin, Michigan Turkey’s vice president of technical services.

In addition, the effectiveness of masks is reliant on a tight seal over the mouth and noise to ensure proper filtration, which can change minute to minute during a workday. Masks can also fog up safety goggles, and they have to be removed for workers to eat. Taza Aya’s technology avoids all of those problems.

As a researcher at U-M, Clack spent years exploring the use of nonthermal plasma to protect livestock. With the arrival of COVID-19 in early 2020, he quickly pivoted to how the technology might be used for personal protection from airborne pathogens.

In October of that year, Taza Aya was named an awardee in the Invisible Shield QuickFire Challenge—a competition created by Johnson & Johnson Innovation in cooperation with the U.S. Department of Health and Human Services. The program sought to encourage the development of technologies that could protect people from airborne viruses while having a minimal impact on daily life.

“We are pleased with the study results as we embark on this journey,” said Alberto Elli, Taza Aya’s CEO. “This real-world product and user testing experience will help us successfully launch the Worker Wearable in 2025.”

Clack and the University of Michigan have a financial interest in Taza Aya.

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sarcozona
35 minutes ago
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I wonder if this is actually more cost effective or comfortable than masks or general indoor air quality improvements.
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So many Americans died from COVID, it boosted Social Security to the tune of $205 billion
Thursday February 20th, 2025 at 1:39 PM

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As the U.S. approaches the fifth anniversary of the official start of the COVID-19 pandemic, new research finds so many Americans died from the virus that the nation’s Social Security trust fund will see a net increase of hundreds of billions of dollars as a result of retirement benefits that will not be paid out.

The working paper from the National Bureau of Economic Research finds that approximately 1.7 million excess deaths among Americans 25 and older occurred between 2020 and 2023 related to the pandemic. Premature deaths related to COVID mean Social Security will not make retirement payments to those individuals in the future, reducing payments by about $294 billion, the researchers found.

At the same time, some of that gain is offset by the lost tax revenue from those individuals, as well as increased survivor benefits to spouses and children of the deceased, resulting in an estimated $205 billion less in future outlays.

Many of the people who died were seniors older than 65 who were already drawing retirement benefits or would soon, and had paid into the system for many years. While most of the excess deaths were white Americans, Black and Hispanic populations saw the highest excess deaths per capita, as did those with lower educational attainment.

Read more from Fortune

Of course, while excess death is one measure of how COVID continues to impact Social Security, there are other ways that the study notes it does not account for—long COVID survivors, for example, are more likely to drop out of the workforce, which could lead to paying less into Social Security over time and possibly needing to tap the safety net’s disability benefits.

“Our analysis suggests a slight improvement in Social Security’s financial health due to excess deaths, driven primarily by the premature death of people who would have received retirement benefits,” the report reads.

The authors—including research scientists from the University of Southern California and an economist from the University of Illinois Urbana-Champaign—note that the savings are “modest.” Social Security is expected to pay out $1.6 trillion in benefits this year, meaning $205 billion is a couple months’ worth of payments.

Did your workplace make our list of the World's Most Admired Companies? Explore this year's list.

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sarcozona
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So many Americans died from Covid, it boosted Social Security by $205…
Thursday February 20th, 2025 at 1:39 PM

1 Share

As the U.S. approaches the fifth anniversary of the official start of the COVID-19 pandemic, new research finds so many Americans died from the virus that the nation’s Social Security trust fund will see a net increase of hundreds of billions of dollars due to retirement benefits that will not be paid out.

The working paper from the National Bureau of Economic Research finds that approximately 1.7 million excess deaths among Americans 25 and older occurred between 2020 and 2023 related to the pandemic. Premature deaths related to Covid mean Social Security will not make retirement payments to those individuals in the future, reducing payments by about $294 billion, the researchers found.

At the same time, some of that gain is offset by the lost tax revenue from those individuals, as well as increased survivor benefits to spouses and children of the deceased, resulting in an estimated $205 billion less in future outlays.

Many of the people who died were seniors older than 65 who were already drawing retirement benefits or would soon, and had paid into the system for many years. While most of the excess deaths were White Americans, Black and Hispanic populations saw the highest excess deaths per capita, as did those with lower educational attainment.

Of course, while excess deaths is one measure of how Covid continues to impact Social Security, there are other ways that the study notes it does not account for—long Covid survivors, for example, are more likely to drop out of the workforce, which could lead to paying less into Social Security over time and possibly needing to tap the safety net’s disability benefits.

“Our analysis suggests a slight improvement in Social Security’s financial health due to excess deaths,
driven primarily by the premature death of people who would have received retirement benefits,” the report reads.

The authors—including research scientists from the University of Southern California and an economist from the University of Illinois Urbana-Champaign—note that the savings are “modest.” This year, Social Security is expected to pay out $1.6 trillion in benefits this year, meaning $205 billion is a couple months’ worth of payments.

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sarcozona
39 minutes ago
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New Analysis Reveals Many Excess Deaths Attributed to Natural Causes Are Actually Uncounted COVID-19 Deaths
Thursday February 20th, 2025 at 1:32 PM

Comments on: New Analysis Reveals Many Excess Deaths Attributed to Natural Causes Are Actually Uncounted COVID-19 Deaths
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sarcozona
46 minutes ago
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If the primary explanation for these deaths were healthcare interruptions and delays in care, the non-COVID excess deaths would likely occur after a peak in reported COVID-19 deaths and subsequent interruptions in care, says study lead author Eugenio Paglino, a PhD student studying demography and sociology at UPenn. “However, this pattern was not observed nationally or in any of the geographic subregions we assessed,” Paglino says.

This is so important because a lot of folks who acknowledge excess deaths are high blame it on the disruptions to healthcare around the start of the pandemic, on anxious patients not coming in until their cancer or whatever was too bad to do anything about. And while certainly those things happened, it did not happen at levels that explain the enormous mortality we've seen.
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Immunological and Antigenic Signatures Associated with Chronic Illnesses after COVID-19 Vaccination | medRxiv
Thursday February 20th, 2025 at 12:09 PM

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sarcozona
2 hours ago
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Glad they’re figuring out the mechanisms here
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