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Article: Assay Techniques and Test Development for COVID-19 Diagnosis.

Carter, Linda J / Garner, Linda V / Smoot, Jeffrey W / Li, Yingzhu / Zhou, Qiongqiong / Saveson, Catherine J / Sasso, Janet M / Gregg, Anne C / Soares, Divya J / Beskid, Tiffany R / Jervey, Susan R / Liu, Cynthia

ACS central science

2020 Volume 6, Issue 5, Page(s) 591–605

Keywords	covid19
Language	English
Publishing date	2020-04-30
Publishing country	United States
Document type	News
ISSN	2374-7943
ISSN	2374-7943
DOI	10.1021/acscentsci.0c00501
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Article: Assay Techniques and Test Development for COVID-19 Diagnosis

Carter, Linda J. / Garner, Linda V. / Smoot, Jeffrey W. / Li, Yingzhu / Zhou, Qiongqiong / Saveson, Catherine J. / Sasso, Janet M. / Gregg, Anne C. / Soares, Divya J. / Beskid, Tiffany R. / Jervey, Susan R. / Liu, Cynthia

ACS Cent. Sci.

Abstract: ... This report describes various assay techniques and tests for COVID-19 diagnosis. Most tests for early ... An ongoing theme of the COVID-19 pandemic is the need for widespread availability of accurate and ... ﬂow immunoassay. This report also provides an overview of current development in COVID-19 ...

Abstract	An ongoing theme of the COVID-19 pandemic is the need for widespread availability of accurate and eﬃcient diagnostic testing for detection of SARS-CoV-2 and antiviral antibodies in infected individuals. This report describes various assay techniques and tests for COVID-19 diagnosis. Most tests for early detection of SARS-CoV-2 RNA rely on the reverse transcription-polymerase chain reaction, but isothermal nucleic acid ampliﬁcation assays, including transcription-mediated ampliﬁcation and CRISPR-based methodologies, are promising alternatives. Identiﬁcation of individuals who have developed antibodies to the SARS-CoV-2 virus requires serological tests, including enzyme-linked immunosorbent assay (ELISA) and lateral ﬂow immunoassay. This report also provides an overview of current development in COVID-19 diagnostic techniques and products to facilitate future improvement and innovation.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #142071
Database	COVID19

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Article ; Online: Assay Techniques and Test Development for COVID-19 Diagnosis

Linda J. Carter / Linda V. Garner / Jeffrey W. Smoot / Yingzhu Li / Qiongqiong Zhou / Catherine J. Saveson / Janet M. Sasso / Anne C. Gregg / Divya J. Soares / Tiffany R. Beskid / Susan R. Jervey / Cynthia Liu

ACS Central Science, Vol 6, Iss 5, Pp 591-

2020 Volume 605

Keywords	Chemistry ; QD1-999
Language	English
Publishing date	2020-04-01T00:00:00Z
Publisher	American Chemical Society
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Assay Techniques and Test Development for COVID-19 Diagnosis

ACS Central Science

2020 Volume 6, Issue 5, Page(s) 591–605

Keywords	covid19
Language	English
Publisher	American Chemical Society (ACS)
Publishing country	us
Document type	Article ; Online
ISSN	2374-7943
DOI	10.1021/acscentsci.0c00501
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Development of an immunofluorescence assay for detection of SARS-CoV-2.

Atiya-Nasagi, Yafit / Milrot, Elad / Makdasi, Efi / Schuster, Ofir / Shmaya, Shlomo / Simon, Irit / Ben-Shmuel, Amir / Beth-Din, Adi / Weiss, Shay / Laskar, Orly

Archives of virology

2022 Volume 167, Issue 4, Page(s) 1041–1049

Abstract: ... 2 at defined concentrations, and infection was monitored at different time points. This test was ... SARS-CoV-2, the etiologic agent of the COVID-19 pandemic, emerged as the cause of a global crisis ... assay is considered highly specific and sensitive, this method cannot determine the infectivity ...

Abstract	SARS-CoV-2, the etiologic agent of the COVID-19 pandemic, emerged as the cause of a global crisis in 2019. Currently, the main method for identification of SARS-CoV-2 is a reverse transcription (RT)-PCR assay designed to detect viral RNA in oropharyngeal (OP) or nasopharyngeal (NP) samples. While the PCR assay is considered highly specific and sensitive, this method cannot determine the infectivity of the sample, which may assist in evaluation of virus transmissibility from patients and breaking transmission chains. Thus, cell-culture-based approaches such as cytopathic effect (CPE) assays are routinely employed for the identification of infectious viruses in NP/OP samples. Despite their high sensitivity, CPE assays take several days and require additional diagnostic tests in order to verify the identity of the pathogen. We have therefore developed a rapid immunofluorescence assay (IFA) for the specific detection of SARS-CoV-2 in NP/OP samples following cell culture infection. Initially, IFA was carried out on Vero E6 cultures infected with SARS-CoV-2 at defined concentrations, and infection was monitored at different time points. This test was able to yield positive signals in cultures infected with 10 pfu/ml at 12 hours postinfection (PI). Increasing the incubation time to 24 hours reduced the detectable infective dose to 1 pfu/ml. These IFA signals occur before the development of CPE. When compared to the CPE test, IFA has the advantages of specificity, rapid detection, and sensitivity, as demonstrated in this work.
MeSH term(s)	COVID-19/diagnosis ; Fluorescent Antibody Technique ; Humans ; Nasopharynx ; Pandemics ; RNA, Viral/genetics ; SARS-CoV-2 ; Sensitivity and Specificity
Chemical Substances	RNA, Viral
Language	English
Publishing date	2022-02-22
Publishing country	Austria
Document type	Journal Article
ZDB-ID	7491-3
ISSN	1432-8798 ; 0304-8608
ISSN (online)	1432-8798
ISSN	0304-8608
DOI	10.1007/s00705-022-05392-z
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Article: CDetection.v2: One-pot assay for the detection of SARS-CoV-2.

Wang, Xinge / Chen, Yangcan / Cheng, Xuejia / Wang, Si-Qi / Hu, Yanping / Feng, Yingmei / Jin, Ronghua / Zhou, Kangping / Liu, Ti / Wang, Jianxing / Pan, Kai / Liu, Bing / Xiang, Jie / Wang, Yanping / Zhou, Qi / Zhang, Ying / Pan, Weiye / Li, Wei

Frontiers in microbiology

2023 Volume 14, Page(s) 1158163

Abstract: ... identification of SARS-CoV-2 and its variants plays a critical role in COVID-19 prevention and control. Currently ... Introduction: The ongoing 2019 coronavirus disease pandemic (COVID-19), caused ... by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and its variants, is a global public health threat. Early diagnosis and ...

Abstract	Introduction: The ongoing 2019 coronavirus disease pandemic (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and its variants, is a global public health threat. Early diagnosis and identification of SARS-CoV-2 and its variants plays a critical role in COVID-19 prevention and control. Currently, the most widely used technique to detect SARS-CoV-2 is quantitative reverse transcription real-time quantitative PCR (RT-qPCR), which takes nearly 1 hour and should be performed by experienced personnel to ensure the accuracy of results. Therefore, the development of a nucleic acid detection kit with higher sensitivity, faster detection and greater accuracy is important. Methods: Here, we optimized the system components and reaction conditions of our previous detection approach by using RT-RAA and Cas12b. Results: We developed a Cas12b-assisted one-pot detection platform (CDetection.v2) that allows rapid detection of SARS-CoV-2 in 30 minutes. This platform was able to detect up to 5,000 copies/ml of SARS-CoV-2 without cross-reactivity with other viruses. Moreover, the sensitivity of this CRISPR system was comparable to that of RT-qPCR when tested on 120 clinical samples. Discussion: The CDetection.v2 provides a novel one-pot detection approach based on the integration of RT-RAA and CRISPR/Cas12b for detecting SARS-CoV-2 and screening of large-scale clinical samples, offering a more efficient strategy for detecting various types of viruses.
Language	English
Publishing date	2023-03-23
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2023.1158163
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Article ; Online: Development and evaluation of a lyophilization protocol for colorimetric RT-LAMP diagnostic assay for COVID-19.

Prado, Nayra Oliveira / Marin, Anelis Maria / Lalli, Larissa Araujo / Sanchuki, Heloisa Bruna Soligo / Wosniaki, Denise Kusma / Nardin, Jeanine Marie / Morales, Hugo Manoel Paz / Blanes, Lucas / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega

Scientific reports

2024 Volume 14, Issue 1, Page(s) 10612

Abstract: ... of a lyophilized colorimetric RT-LAMP COVID-19 detection assay with diagnostic sensitivity and ... transport and storage. We conducted tests to identify the ideal excipients that maintain the molecular ... Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional ...

Abstract	Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional tools. During the 2020 global health crisis, efforts intensified to optimize the production and delivery of molecular diagnostic kits for detecting SARS-CoV-2. During this period, RT-LAMP emerged as a significant focus. However, the thermolability of the reagents used in this technique necessitates special low-temperature infrastructure for transport, storage, and conservation. These requirements limit distribution capacity and necessitate cost-increasing adaptations. Consequently, this report details the development of a lyophilization protocol for reagents in a colorimetric RT-LAMP diagnostic kit to detect SARS-CoV-2, facilitating room-temperature transport and storage. We conducted tests to identify the ideal excipients that maintain the molecular integrity of the reagents and ensure their stability during room-temperature storage and transport. The optimal condition identified involved adding 5% PEG 8000 and 75 mM trehalose to the RT-LAMP reaction, which enabled stability at room temperature for up to 28 days and yielded an analytical and diagnostic sensitivity and specificity of 83.33% and 90%, respectively, for detecting SARS-CoV-2. This study presents the results of a lyophilized colorimetric RT-LAMP COVID-19 detection assay with diagnostic sensitivity and specificity comparable to RT-qPCR, particularly in samples with high viral load.
MeSH term(s)	Freeze Drying ; Humans ; COVID-19/diagnosis ; COVID-19/virology ; SARS-CoV-2/isolation & purification ; SARS-CoV-2/genetics ; Colorimetry/methods ; Nucleic Acid Amplification Techniques/methods ; Molecular Diagnostic Techniques/methods ; RNA, Viral/analysis ; RNA, Viral/genetics ; Sensitivity and Specificity ; Reagent Kits, Diagnostic/standards ; COVID-19 Nucleic Acid Testing/methods
Chemical Substances	RNA, Viral ; Reagent Kits, Diagnostic
Language	English
Publishing date	2024-05-09
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-61163-7
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Article ; Online: Lateral Flow Assay Biotesting by Utilizing Plasmonic Nanoparticles Made of Inexpensive Metals─Replacing Colloidal Gold.

Bahamondes Lorca, Veronica A / Ávalos-Ovando, Oscar / Sikeler, Christoph / Ijäs, Heini / Santiago, Eva Yazmin / Skelton, Eli / Wang, Yong / Yang, Ruiqi / Cimatu, Katherine Leslee Asetre / Baturina, Olga / Wang, Zhewei / Liu, Jundong / Slocik, Joseph M / Wu, Shiyong / Ma, Dongling / Pastukhov, Andrei / Kabashin, Andrei V / Kordesch, Martin E / Govorov, Alexander O

Nano letters

2024 Volume 24, Issue 20, Page(s) 6069–6077

Abstract: ... to detect target analytes in biological samples. This proven concept was primarily used during the COVID-19 ... in commercial testing kits is the colloidal synthesis, our development with the Cu@Au and the laser-ablation ... pandemic with gold-NP-based lateral flow assays (LFAs). Considering the gold price and its worldwide ...

Abstract	Nanoparticles (NPs) can be conjugated with diverse biomolecules and employed in biosensing to detect target analytes in biological samples. This proven concept was primarily used during the COVID-19 pandemic with gold-NP-based lateral flow assays (LFAs). Considering the gold price and its worldwide depletion, here we show that novel plasmonic NPs based on inexpensive metals, titanium nitride (TiN) and copper covered with a gold shell (Cu@Au), perform comparable to or even better than gold nanoparticles. After conjugation, these novel nanoparticles provided high figures of merit for LFA testing, such as high signals and specificity and robust naked-eye signal recognition. Since the main cost of Au NPs in commercial testing kits is the colloidal synthesis, our development with the Cu@Au and the laser-ablation-fabricated TiN NPs is exciting, offering potentially inexpensive plasmonic nanomaterials for various bioapplications. Moreover, our machine learning study showed that biodetection with TiN is more accurate than that with Au.
MeSH term(s)	Metal Nanoparticles/chemistry ; Titanium/chemistry ; Gold/chemistry ; Copper/chemistry ; Biosensing Techniques/methods ; Biosensing Techniques/economics ; Humans ; COVID-19/virology ; COVID-19/diagnosis ; Gold Colloid/chemistry ; SARS-CoV-2/isolation & purification
Chemical Substances	titanium nitride
Language	English
Publishing date	2024-05-13
Publishing country	United States
Document type	Journal Article
ISSN	1530-6992
ISSN (online)	1530-6992
DOI	10.1021/acs.nanolett.4c01022
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Article ; Online: Targets and assay types for COVID-19 diagnosis.

Bostan, Marinela / Ataman, Marius / Bostan, Ioana Stefania / Bleotu, Coralia

Journal of immunoassay & immunochemistry

2021 Volume 41, Issue 6, Page(s) 946–959

Abstract: ... the specific targets and the sensitivity of each assay used for COVID-19 diagnosis can lead to more efficient ... this technique in all clinical laboratories limits its widespread use. In the case of COVID-19, these tests can be used for the triage ... such as reverse transcription PCR (RT-PCR) to diagnose Coronavirus 2019 disease (COVID-19). Although remarkable progress has ...

Abstract	The lack of complete information on the immune response dynamics to infection with severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) has led to the use of mainly molecular tests such as reverse transcription PCR (RT-PCR) to diagnose Coronavirus 2019 disease (COVID-19). Although remarkable progress has been made in developing effective RT-PCR kits, the lack of specific equipment required to perform this technique in all clinical laboratories limits its widespread use. In the case of COVID-19, these tests can be used for the triage of symptomatic patients, for testing the contacts of confirmed cases, and also for the analysis and monitoring of the situation. Along with molecular tests involving reverse transcription PCR, various laboratory tests can identify the specific anti-viral antibodies or viral antigens. This review seeks to describe the targets and diagnostic methods available or currently in development for SARS-CoV-2 infection, including reverse transcription PCR (RT-PCR), serologic immunoassays (SIA) and the protein microarray method (PMM). Knowing the specific targets and the sensitivity of each assay used for COVID-19 diagnosis can lead to more efficient detection of infected patients and it can provide better management of the pandemic status.
Language	English
Publishing date	2021-02-04
Publishing country	England
Document type	Journal Article
ZDB-ID	2050610-7
ISSN	1532-4230 ; 1532-1819
ISSN (online)	1532-4230
ISSN	1532-1819
DOI	10.1080/15321819.2020.1862866
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Article: Development of a RT-LAMP assay for detection of SARS-CoV-2.

Nandi, Shyam Sundar / Lambe, Upendra Pradeep / Sawant, Sonali Ankush / Gohil, Trupti / Deshpande, Jagadish

The Indian journal of medical research

2022 Volume 155, Issue 1, Page(s) 148–155

Abstract: ... assay primers were designed to detect envelop (E) and nucleocapsid (N) genes of SARS-CoV-2. Positive ... by comparing the results obtained with a commercial rRT-PCR kit.: Results: The RT-LAMP assay for E and N ... sensitivity and specificity of the LAMP assay were 98.46 and 100 per cent, respectively, as compared to the rRT-PCR ...

Abstract	Background & objectives: The pandemic of SARS-COV-2 began in Wuhan, China in December 2019 and has caused more than 101 million cases worldwide. Diagnostic technologies possessing sensitivity and specificity equivalent to real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) assays are needed to ramp up testing capacity in most countries. Newer platforms need to be technically less demanding, require minimum equipment and reduce turn-around time for reporting results. The objective of this study was to exploit loop-mediated isothermal amplification (LAMP) for the detection of SARS-CoV-2 and evaluate its performance by comparison with rRT-PCR. Methods: Reverse-transcription LAMP (RT-LAMP) assay primers were designed to detect envelop (E) and nucleocapsid (N) genes of SARS-CoV-2. Positive control RNA was prepared by in vitro transcription of E and N genes clones. RT-LAMP amplification reactions were incubated at 65°C for 30 min. Results were recorded visually. RT-LAMP results were evaluated by comparing the results obtained with a commercial rRT-PCR kit. Results: The RT-LAMP assay for E and N genes was carried out in separate tubes. RT-LAMP detected about 40 copies of SARS-CoV-2 RNA per reaction. A total of 253 throat swabs were tested using the RT-LAMP assay. The overall diagnostic sensitivity and specificity of the LAMP assay were 98.46 and 100 per cent, respectively, as compared to the rRT-PCR. Interpretation & conclusions: SARS-CoV-2 RT-LAMP assay was designed, standardized and evaluated. The assay showed diagnostic sensitivity and specificity equivalent to rRT-PCR assays. The assay will be useful to increase testing capacity for the detection of SARS-CoV-2 in the country.
MeSH term(s)	COVID-19/diagnosis ; COVID-19 Testing ; Humans ; Molecular Diagnostic Techniques ; Nucleic Acid Amplification Techniques/methods ; RNA, Viral/genetics ; SARS-CoV-2/genetics ; Sensitivity and Specificity
Chemical Substances	RNA, Viral
Language	English
Publishing date	2022-03-10
Publishing country	India
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390883-5
ISSN	0971-5916 ; 0019-5340
ISSN	0971-5916 ; 0019-5340
DOI	10.4103/ijmr.IJMR_713_21
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