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Article ; Online: Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients.

Yu, Fengting / Yan, Liting / Wang, Nan / Yang, Siyuan / Wang, Linghang / Tang, Yunxia / Gao, Guiju / Wang, Sa / Ma, Chengjie / Xie, Ruming / Wang, Fang / Tan, Chianru / Zhu, Lingxiang / Guo, Yong / Zhang, Fujie

Clinical infectious diseases : an official publication of the Infectious Diseases Society of America

2020 Volume 71, Issue 15, Page(s) 793–798

Abstract: ... samples was then compared and the average viral load in sputum (17 429 ± 6920 copies/test) was found to be ... according to ddPCR with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory ... diagnosis and treatment.: Methods: A total of 323 samples from 76 COVID-19-confirmed patients were ...

Abstract	Background: Coronavirus disease 2019 (COVID-19) has become a public health emergency. The widely used reverse transcription-polymerase chain reaction (RT-PCR) method has limitations for clinical diagnosis and treatment. Methods: A total of 323 samples from 76 COVID-19-confirmed patients were analyzed by droplet digital PCR (ddPCR) and RT-PCR based 2 target genes (ORF1ab and N). Nasal swabs, throat swabs, sputum, blood, and urine were collected. Clinical and imaging data were obtained for clinical staging. Results: In 95 samples that tested positive by both methods, the cycle threshold (Ct) of RT-PCR was highly correlated with the copy number of ddPCR (ORF1ab gene, R2 = 0.83; N gene, R2 = 0.87). Four (4/161) negative and 41 (41/67) single-gene positive samples tested by RT-PCR were positive according to ddPCR with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared and the average viral load in sputum (17 429 ± 6920 copies/test) was found to be significantly higher than in throat swabs (2552 ± 1965 copies/test, P < .001) and nasal swabs (651 ± 501 copies/test, P < .001). Furthermore, the viral loads in the early and progressive stages were significantly higher than that in the recovery stage (46 800 ± 17 272 vs 1252 ± 1027, P < .001) analyzed by sputum samples. Conclusions: Quantitative monitoring of viral load in lower respiratory tract samples helps to evaluate disease progression, especially in cases of low viral load.
MeSH term(s)	Adult ; Betacoronavirus/genetics ; COVID-19 ; Coronavirus Infections/diagnosis ; Coronavirus Infections/virology ; Diagnostic Tests, Routine/methods ; False Negative Reactions ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral/diagnosis ; Pneumonia, Viral/virology ; Real-Time Polymerase Chain Reaction/methods ; Respiratory System/virology ; SARS-CoV-2 ; Serologic Tests/methods ; Sputum/virology ; Viral Load/methods
Keywords	covid19
Language	English
Publishing date	2020-03-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1099781-7
ISSN	1537-6591 ; 1058-4838
ISSN (online)	1537-6591
ISSN	1058-4838
DOI	10.1093/cid/ciaa345
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Article ; Online: Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients

Clinical Infectious Diseases

2020 Volume 71, Issue 15, Page(s) 793–798

Abstract: ... and the average viral load in sputum (17 429 ± 6920 copies/test) was found to be significantly higher ... with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared ... diagnosis and treatment. Methods A total of 323 samples from 76 COVID-19–confirmed patients were analyzed ...

Abstract	Abstract Background Coronavirus disease 2019 (COVID-19) has become a public health emergency. The widely used reverse transcription–polymerase chain reaction (RT-PCR) method has limitations for clinical diagnosis and treatment. Methods A total of 323 samples from 76 COVID-19–confirmed patients were analyzed by droplet digital PCR (ddPCR) and RT-PCR based 2 target genes (ORF1ab and N). Nasal swabs, throat swabs, sputum, blood, and urine were collected. Clinical and imaging data were obtained for clinical staging. Results In 95 samples that tested positive by both methods, the cycle threshold (Ct) of RT-PCR was highly correlated with the copy number of ddPCR (ORF1ab gene, R2 = 0.83; N gene, R2 = 0.87). Four (4/161) negative and 41 (41/67) single-gene positive samples tested by RT-PCR were positive according to ddPCR with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared and the average viral load in sputum (17 429 ± 6920 copies/test) was found to be significantly higher than in throat swabs (2552 ± 1965 copies/test, P < .001) and nasal swabs (651 ± 501 copies/test, P < .001). Furthermore, the viral loads in the early and progressive stages were significantly higher than that in the recovery stage (46 800 ± 17 272 vs 1252 ± 1027, P < .001) analyzed by sputum samples. Conclusions Quantitative monitoring of viral load in lower respiratory tract samples helps to evaluate disease progression, especially in cases of low viral load.
Keywords	Microbiology (medical) ; Infectious Diseases ; covid19
Language	English
Publisher	Oxford University Press (OUP)
Publishing country	uk
Document type	Article ; Online
ZDB-ID	1099781-7
ISSN	1537-6591 ; 1058-4838
ISSN (online)	1537-6591
ISSN	1058-4838
DOI	10.1093/cid/ciaa345
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Article: Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients

Clin Infect Dis

Abstract: ... and the average viral load in sputum (17â 429â ±â 6920 copies/test) was found to be significantly ... with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared ... diagnosis and treatment. METHODS: A total of 323 samples from 76 COVID-19-confirmed patients were analyzed ...

Abstract	BACKGROUND: Coronavirus disease 2019 (COVID-19) has become a public health emergency. The widely used reverse transcription-polymerase chain reaction (RT-PCR) method has limitations for clinical diagnosis and treatment. METHODS: A total of 323 samples from 76 COVID-19-confirmed patients were analyzed by droplet digital PCR (ddPCR) and RT-PCR based 2 target genes (ORF1ab and N). Nasal swabs, throat swabs, sputum, blood, and urine were collected. Clinical and imaging data were obtained for clinical staging. RESULTS: In 95 samples that tested positive by both methods, the cycle threshold (Ct) of RT-PCR was highly correlated with the copy number of ddPCR (ORF1ab gene, R2â =â 0.83; N gene, R2â =â 0.87). Four (4/161) negative and 41 (41/67) single-gene positive samples tested by RT-PCR were positive according to ddPCR with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared and the average viral load in sputum (17â 429â ±â 6920 copies/test) was found to be significantly higher than in throat swabs (2552â ±â 1965 copies/test, Pâ <â .001) and nasal swabs (651â ±â 501 copies/test, Pâ <â .001). Furthermore, the viral loads in the early and progressive stages were significantly higher than that in the recovery stage (46â 800â ±â 17â 272 vs 1252â ±â 1027, Pâ <â .001) analyzed by sputum samples. CONCLUSIONS: Quantitative monitoring of viral load in lower respiratory tract samples helps to evaluate disease progression, especially in cases of low viral load.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #17963
Database	COVID19

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Article ; Online: Quantitative SARS-CoV-2 subgenomic RNA as a surrogate marker for viral infectivity: Comparison between culture isolation and direct sgRNA quantification.

Scutari, Rossana / Renica, Silvia / Cento, Valeria / Nava, Alice / Sammartino, Josè Camilla / Ferrari, Alessandro / Pani, Arianna / Merli, Marco / Fanti, Diana / Vismara, Chiara / Scaglione, Francesco / Puoti, Massimo / Bandera, Alessandra / Gori, Andrea / Piralla, Antonio / Baldanti, Fausto / Perno, Carlo Federico / Alteri, Claudia

PloS one

2023 Volume 18, Issue 9, Page(s) e0291120

Abstract: ... swabs collected from SARS-CoV-2 positive hospitalized patients. Among the 51 samples, 14 were SARS-CoV-2 ... patients with sgRNA equal to or higher than sgRNA cut-offs. Overall, this study suggests that SARS-CoV-2 ... of this assay in available SARS-CoV-2 diagnostics procedure might help in optimizing fragile patients monitoring ...

Abstract	Detection of subgenomic (sg) SARS-CoV-2 RNAs are frequently used as a correlate of viral infectiousness, but few data about correlation between sg load and viable virus are available. Here, we defined concordance between culture isolation and E and N sgRNA quantification by ddPCR assays in 51 nasopharyngeal swabs collected from SARS-CoV-2 positive hospitalized patients. Among the 51 samples, 14 were SARS-CoV-2 culture-positive and 37 were negative. According to culture results, the sensitivity and specificity of E and N sgRNA assays were 100% and 100%, and 84% and 86%, respectively. ROC analysis showed that the best E and N cut-offs to predict positive culture isolation were 32 and 161 copies/mL respectively, with an AUC (95% CI) of 0.96 (0.91-1.00) and 0.96 (0.92-1.00), and a diagnostic accuracy of 88% and 92%, respectively. Even if no significant correlations were observed between sgRNA amount and clinical presentation, a higher number of moderate/severe cases and lower number of days from symptoms onset characterized patients with sgRNA equal to or higher than sgRNA cut-offs. Overall, this study suggests that SARS-CoV-2 sgRNA quantification could be helpful to estimate the replicative activity of SARS-CoV-2 and can represent a valid surrogate marker to efficiently recognize patients with active infection. The inclusion of this assay in available SARS-CoV-2 diagnostics procedure might help in optimizing fragile patients monitoring and management.
MeSH term(s)	Humans ; COVID-19/diagnosis ; SARS-CoV-2/genetics ; Subgenomic RNA ; Virus Diseases ; Biomarkers ; RNA
Chemical Substances	Subgenomic RNA ; Biomarkers ; RNA (63231-63-0)
Language	English
Publishing date	2023-09-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0291120
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Article ; Online: Retrospective quantitative detection of SARS-CoV-2 by digital PCR showing high accuracy for low viral load specimens.

Lu, Renfei / Wang, Jian / Li, Min / He, Jing / Wang, Yaqi / Dong, Jia / Cai, Weihua

Journal of infection in developing countries

2022 Volume 16, Issue 1, Page(s) 10–15

Abstract: ... the infection status and viral load of severe acute respiratory syndrome coronavirus 2 in Nantong city, China ... A total of 103 clinical specimens from 31 patients were collected and tested by digital PCR and ... than reverse-transcription PCR, especially for low viral load specimens. ...

Abstract	Introduction: Accurate detection of severe acute respiratory syndrome coronavirus 2 is critical for diagnosis and disease status evaluation of Coronavirus disease 2019. We retrospectively evaluated the infection status and viral load of severe acute respiratory syndrome coronavirus 2 in Nantong city, China, using a quantitative digital polymerase chain reaction and reverse-transcription PCR. Methodology: A total of 103 clinical specimens from 31 patients were collected and tested by digital PCR and reverse-transcription PCR. Results: The overall accuracy of digital PCR was 96.8%, which was higher than the overall accuracy of 87.1% for reverse-transcription PCR. 4 (3.88%) specimens for ORF1ab and 22 (21.36%) specimens for N gene were negative by reverse-transcription PCR but positive by digital PCR. 3 (2.91%, 3/103) specimens of ORF1ab were positive by reverse-transcription PCR but negative by digital PCR. The digital PCR assay exhibited higher sensitivity to measure the N gene than the ORF1ab gene (p < 0.01). Conclusions: Our results showed that digital PCR assay provides more reliable detection of Coronavirus disease 2019 than reverse-transcription PCR, especially for low viral load specimens.
MeSH term(s)	COVID-19 ; Humans ; Polymerase Chain Reaction ; RNA, Viral/analysis ; RNA, Viral/genetics ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2 ; Sensitivity and Specificity ; Viral Load
Chemical Substances	RNA, Viral
Language	English
Publishing date	2022-01-31
Publishing country	Italy
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2394024-4
ISSN	1972-2680 ; 2036-6590
ISSN (online)	1972-2680
ISSN	2036-6590
DOI	10.3855/jidc.15315
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Article ; Online: Quantitative analysis of different respiratory specimens on two automated test systems for detection of SARS-CoV-2 RNA.

Lübke, Nadine / Repges, Katharina / Menne, Christopher / Walker, Andreas / Jensen, Björn-Erik O / Freise, Noemi F / Gliga, Smaranda / Eickhoff, Simon B / Bosse, Hans Martin / Adams, Ortwin / Timm, Jörg

Diagnostic microbiology and infectious disease

2022 Volume 105, Issue 1, Page(s) 115800

Abstract: ... showed reliable detection and quantification of SARS-CoV-2 RNA, with nasopharyngeal swabs showing ... quantification of SARS-CoV-2 was performed on cobas®6800 (Roche) and NeuMoDx™ (Qiagen) systems. Both assays ... and tested simultaneously from a total of 36 hospitalized symptomatic COVID-19 patients. Detection and ...

Abstract	Molecular testing of SARS-CoV-2 RNA is essential during the pandemic. Here, we compared the results of different respiratory specimens including anterior nasal swabs, pharyngeal swabs, saliva swabs, and gargle lavage samples to nasopharyngeal swabs on two automated SARS-CoV-2 test systems. Samples were collected and tested simultaneously from a total of 36 hospitalized symptomatic COVID-19 patients. Detection and quantification of SARS-CoV-2 was performed on cobas®6800 (Roche) and NeuMoDx™ (Qiagen) systems. Both assays showed reliable detection and quantification of SARS-CoV-2 RNA, with nasopharyngeal swabs showing the highest sensitivity. SARS-CoV-2 RNA concentrations in other respiratory specimens were lower (mean 2.5 log10 copies/ml) or even undetectable in up to 20%. These data clearly indicate that not all respiratory materials are equally suitable for the management of hospitalized patients, especially, in the late phase of COVID-19, when the viral phase subsides and inflammation becomes the predominant factor, making detection of even lower viral loads increasingly important.
MeSH term(s)	Humans ; SARS-CoV-2/genetics ; COVID-19/diagnosis ; RNA, Viral/genetics ; Pandemics ; COVID-19 Testing ; Saliva ; Nasopharynx ; Specimen Handling/methods
Chemical Substances	RNA, Viral
Language	English
Publishing date	2022-08-26
Publishing country	United States
Document type	Journal Article
ZDB-ID	604920-5
ISSN	1879-0070 ; 0732-8893
ISSN (online)	1879-0070
ISSN	0732-8893
DOI	10.1016/j.diagmicrobio.2022.115800
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Article ; Online: Comparison of qualitative and quantitative analyses of COVID-19 clinical samples.

Dang, Yan / Liu, Ning / Tan, Chianru / Feng, Yingmei / Yuan, Xingxing / Fan, Dongdong / Peng, Yanke / Jin, Ronghua / Guo, Yong / Lou, Jinli

Clinica chimica acta; international journal of clinical chemistry

2020 Volume 510, Page(s) 613–616

Abstract: Background: Qualitative and quantitative detection of nucleic acids of SARS-CoV-2, the pathogen ... patients without COVID-19 were collected. Reverse transcriptase quantitative PCR (RT-qPCR) and droplet ... prevention, and control.: Methods: A total of 117 samples from 30 patients with confirmed COVID-19 and 61 ...

Abstract	Background: Qualitative and quantitative detection of nucleic acids of SARS-CoV-2, the pathogen that causes coronavirus disease 2019 (COVID-19), plays a significant role in COVID-19 diagnosis, surveillance, prevention, and control. Methods: A total of 117 samples from 30 patients with confirmed COVID-19 and 61 patients without COVID-19 were collected. Reverse transcriptase quantitative PCR (RT-qPCR) and droplet digital PCR (ddPCR) were used for qualitative and quantitative analyses of these samples to evaluate the diagnostic performance and applicability of the two methods. Results: The positive detection rates of RT-qPCR and ddPCR were 93.3% and 100%, respectively. Among the 117 samples, 6 samples were tested single-gene positive by RT-qPCR but positive by ddPCR, and 3 samples were tested negative by RT-qPCR but positive by ddPCR. The viral load of samples with inconsistent results were relatively low (3.1-20.5 copies/test). There were 17 samples (37%) with a viral load below 20 copies/test among the 46 positive samples, and only 9 of them were successfully detected by RT-qPCR. A severe patient was dynamically monitored. All 6 samples from this patient were tested negative by RT-qPCR, but 4 samples were tested positive by ddPCR with a low viral load. Conclusion: Qualitative analysis of COVID-19 samples can meet the needs of clinical screening and diagnosis, while quantitative analysis provides more information to the research community. Although both ddPCR and RT-qPCR can provide qualitative and quantitative results, ddPCR showed higher sensitivity and lower limit of detection than RT-qPCR, and it does not rely on the standard curve to quantify viral load. Therefore, ddPCR offers greater advantages than RT-qPCR.
MeSH term(s)	Adult ; Aged ; COVID-19 ; Case-Control Studies ; Coronavirus Infections/diagnosis ; Coronavirus Infections/genetics ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral/diagnosis ; Pneumonia, Viral/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Viral Load
Keywords	covid19
Language	English
Publishing date	2020-08-25
Publishing country	Netherlands
Document type	Comparative Study ; Journal Article
ZDB-ID	80228-1
ISSN	1873-3492 ; 0009-8981
ISSN (online)	1873-3492
ISSN	0009-8981
DOI	10.1016/j.cca.2020.08.033
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Article ; Online: Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix.

Queiroz, Jackson Alves da Silva / Rampazzo, Rita de Cássia Pontello / Filho, Edivá Basílio da Silva / Oliveira, Gabriella Sgorlon / Oliveira, Suyane da Costa / Souza, Luan Felipo Botelho / Pereira, Soraya Dos Santos / Rodrigues, Moreno Magalhães de Souza / Maia, Adriana Cristina Salvador / da Silva, Cicileia Correia / Mendonça, Aline Linhares Ferreira de Melo / Lugtenburg, Celina Aparecida Bertoni / Aguiar, Francisco de Assis Araújo / Rodrigues, Rosiane de Souza Soares / Santos, Caio Henrique Nemeth / Guimarães, Alice Paula Di Sabatino / Máximo, Fernando Rodrigues / Santos, Alcione de Oliveira Dos / Krieger, Marco Aurélio /

Salcedo, Juan Miguel Villalobos / Dall'Acqua, Deusilene Souza Vieira

International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases

2021 Volume 104, Page(s) 373–378

Abstract: ... by Severe Acute Respiratory Syndrome Virus 2 (SARS-CoV-2) that emerged in China in late 2019. The rapid viral spread has made ... of quantifying as few as 2.5 copies/reaction and an analysis of 244 patients with known results selected by cross ... In this population, it was possible to quantify patients with between 2.59 and 3.5 × 10: Conclusion: This assay ...

Abstract	Introduction: Coronavirus disease-2019 (COVID-19) is a disease caused by Severe Acute Respiratory Syndrome Virus 2 (SARS-CoV-2) that emerged in China in late 2019. The rapid viral spread has made the disease a public health emergency of worldwide concern. The gold standard for diagnosing SARS-CoV-2 is reverse transcription followed by qualitative real-time polymerase chain reaction (RT-qPCR); however, the role of viral load quantification has not been thoroughly investigated yet. Objective: The aim of this study was to develop a high-precision quantitative one-step RT-qPCR reaction using the association of the viral target and the human target in the same reaction. Methods: The assay standardization involved the absolute quantification method, with serial dilutions of a plasmid with the N gene in a biological matrix to build a standard curve. Results and discussion: The results demonstrated the possibility of quantifying as few as 2.5 copies/reaction and an analysis of 244 patients with known results selected by cross-section that revealed 100% agreement with a qualitative RT-qPCR assay registered by Anvisa. In this population, it was possible to quantify patients with between 2.59 and 3.5 × 10 Conclusion: This assay can be a useful tool for a proper patient management, because the level and duration of viral replication are important factors to assess the risk of transmission and to guide decisions regarding the isolation and release of patients; an accurate diagnosis is critical information, whereas the current COVID-19 pandemic represents the biggest current global health problem.
MeSH term(s)	Adolescent ; Adult ; Aged ; Aged, 80 and over ; COVID-19/diagnosis ; COVID-19/virology ; COVID-19 Nucleic Acid Testing/methods ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Middle Aged ; Multiplex Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Reference Standards ; SARS-CoV-2/isolation & purification ; Sensitivity and Specificity ; Viral Load ; Young Adult
Language	English
Publishing date	2021-01-09
Publishing country	Canada
Document type	Journal Article
ZDB-ID	1331197-9
ISSN	1878-3511 ; 1201-9712
ISSN (online)	1878-3511
ISSN	1201-9712
DOI	10.1016/j.ijid.2021.01.001
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Article ; Online: Comparison of automated SARS-CoV-2 antigen test for COVID-19 infection with quantitative RT-PCR using 313 nasopharyngeal swabs, including from seven serially followed patients.

Hirotsu, Yosuke / Maejima, Makoto / Shibusawa, Masahiro / Nagakubo, Yuki / Hosaka, Kazuhiro / Amemiya, Kenji / Sueki, Hitomi / Hayakawa, Miyoko / Mochizuki, Hitoshi / Tsutsui, Toshiharu / Kakizaki, Yumiko / Miyashita, Yoshihiro / Yagi, Shintaro / Kojima, Satoshi / Omata, Masao

International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases

2020 Volume 99, Page(s) 397–402

Abstract: ... can rapidly identify SARS-CoV-2-infected individuals with moderate to high viral loads and may be helpful ... individuals) were analyzed for SARS-CoV-2 with quantitative RT-PCR (RT-qPCR) and then subjected to LUMIPULSE ... samples from 7 infected patients and 231 individual samples from 4 infected patients and 215 uninfected ...

Abstract	In routine clinical practice, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is determined by reverse-transcription PCR (RT-PCR). In the current pandemic, a more rapid and high-throughput method is in growing demand. Here, we validated the performance of a new antigen test (LUMIPULSE) based on chemiluminescence enzyme immunoassay. A total of 313 nasopharyngeal swabs (82 serial samples from 7 infected patients and 231 individual samples from 4 infected patients and 215 uninfected individuals) were analyzed for SARS-CoV-2 with quantitative RT-PCR (RT-qPCR) and then subjected to LUMIPULSE. We determined the cutoff value for antigen detection using receiver operating characteristic curve analysis and compared the performance of the antigen test with that of RT-qPCR. We also compared the viral loads and antigen levels in serial samples from seven infected patients. Using RT-qPCR as the reference, the antigen test exhibited 55.2% sensitivity and 99.6% specificity, with a 91.4% overall agreement rate (286/313). In specimens with > 100 viral copies and between 10 and 100 copies, the antigen test showed 100% and 85% concordance with RT-qPCR, respectively. This concordance declined with lower viral loads. In the serially followed patients, the antigen levels showed a steady decline, along with viral clearance. This gradual decline was in contrast with the abrupt positive-to-negative and negative-to-positive status changes observed with RT-qPCR, particularly in the late phase of infection. In summary, the LUMIPULSE antigen test can rapidly identify SARS-CoV-2-infected individuals with moderate to high viral loads and may be helpful for monitoring viral clearance in hospitalized patients.
MeSH term(s)	Betacoronavirus ; COVID-19 ; COVID-19 Testing ; Clinical Laboratory Techniques/methods ; Coronavirus Infections/diagnosis ; Humans ; Immunoenzyme Techniques ; Luminescent Measurements ; Nasal Cavity/virology ; Pandemics ; Pneumonia, Viral/diagnosis ; Real-Time Polymerase Chain Reaction/methods ; SARS-CoV-2 ; Sensitivity and Specificity ; Viral Load
Keywords	covid19
Language	English
Publishing date	2020-08-12
Publishing country	Canada
Document type	Comparative Study ; Journal Article ; Validation Study
ZDB-ID	1331197-9
ISSN	1878-3511 ; 1201-9712
ISSN (online)	1878-3511
ISSN	1201-9712
DOI	10.1016/j.ijid.2020.08.029
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Article ; Online: Comparison of automated SARS-CoV-2 antigen test for COVID-19 infection with quantitative RT-PCR using 313 nasopharyngeal swabs, including from seven serially followed patients

Yosuke Hirotsu / Makoto Maejima / Masahiro Shibusawa / Yuki Nagakubo / Kazuhiro Hosaka / Kenji Amemiya / Hitomi Sueki / Miyoko Hayakawa / Hitoshi Mochizuki / Toshiharu Tsutsui / Yumiko Kakizaki / Yoshihiro Miyashita / Shintaro Yagi / Satoshi Kojima / Masao Omata

International Journal of Infectious Diseases, Vol 99, Iss , Pp 397-

2020 Volume 402

Abstract	In routine clinical practice, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is determined by reverse-transcription PCR (RT-PCR). In the current pandemic, a more rapid and high-throughput method is in growing demand. Here, we validated the performance of a new antigen test (LUMIPULSE) based on chemiluminescence enzyme immunoassay. A total of 313 nasopharyngeal swabs (82 serial samples from 7 infected patients and 231 individual samples from 4 infected patients and 215 uninfected individuals) were analyzed for SARS-CoV-2 with quantitative RT-PCR (RT-qPCR) and then subjected to LUMIPULSE. We determined the cutoff value for antigen detection using receiver operating characteristic curve analysis and compared the performance of the antigen test with that of RT-qPCR. We also compared the viral loads and antigen levels in serial samples from seven infected patients. Using RT-qPCR as the reference, the antigen test exhibited 55.2% sensitivity and 99.6% specificity, with a 91.4% overall agreement rate (286/313). In specimens with > 100 viral copies and between 10 and 100 copies, the antigen test showed 100% and 85% concordance with RT-qPCR, respectively. This concordance declined with lower viral loads. In the serially followed patients, the antigen levels showed a steady decline, along with viral clearance. This gradual decline was in contrast with the abrupt positive-to-negative and negative-to-positive status changes observed with RT-qPCR, particularly in the late phase of infection. In summary, the LUMIPULSE antigen test can rapidly identify SARS-CoV-2-infected individuals with moderate to high viral loads and may be helpful for monitoring viral clearance in hospitalized patients.
Keywords	SARS-CoV-2 ; COVID-19 ; Antigen ; RT-qPCR ; Infection ; Immunoassay ; Infectious and parasitic diseases ; RC109-216 ; covid19
Subject code	610
Language	English
Publishing date	2020-10-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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