Development and comparison of novel multiple cross displacement amplification (MCDA) assays with other nucleic acid amplification methods for SARS-CoV-2 detection

Luu, Laurence Don Wai; Payne, Michael; Zhang, Xiaomei; Luo, Lijuan; Lan, Ruiting

doi:10.1038/s41598-021-81518-8

Cited by 16 publications

(10 citation statements)

References 17 publications

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“…However, due to some limitations in nucleic acid testing, such as long detection cycle, expensive reagents and high requirements for sampling, and technicians and laboratory conditions, many COVID‐19 cases are not diagnosed accurately and timely. 6 , 7 Up to now, 369 cases of asymptomatic infection in China were still under medical observation, including 352 cases imported from abroad. 8 Therefore, there is an urgent need to explore a rapid, simple, and feasible method for the diagnosis and screening of COVID‐19 patients, especially the asymptomatic cases.…”

Section: Introductionmentioning

confidence: 99%

Dynamic changes of IgM and IgG antibodies in asymptomatic patients as an effective way to detect SARS‐CoV‐2 infection

Shen

Zhu

et al. 2021

Clinical Laboratory Analysis

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Background: COVID-19 has become a global pandemic, and close contacts and asymptomatic patients are worthy of attention.Methods: A total of 1844 people in close contacts with 76 COVID-19 patients were investigated, and nasopharyngeal swabs and venous blood were collected for centralized medical quarantine observation. Real-time fluorescence was used to detect SARS-CoV-2 nucleic acid in nasopharyngeal swabs of all close contacts, and the colloidal gold method was used to detect serum-specific antibodies. Levels of IgM-and IgG-specific antibodies were detected quantitatively through chemiluminescence from the first nucleic acid turned negative date (0 week) and on weekly intervals of ≤1 week, 1-2 weeks, 2-3 weeks, 3-4 weeks, 4-5 weeks, 5-6 weeks, and 6-7 weeks. Results:The total positive rate of the colloidal gold method (88.5%, 23/26) was significantly higher (χ 2 = 59.182, p < 0.001) than that of the healthy control group (2.0%, 1/50). There was significant difference in IgG concentration at different time points (0-7 weeks) after negative nucleic acid conversion (χ 2 = 14.034, p = 0.029). Serum IgG levels were significantly higher at weekly time points of 4-5 weeks (Z = −2.399, p = 0.016), 5-6 weeks (Z = −2.049, p = 0.040), and 6-7 weeks (Z = −2.197, p = 0.028) compared with 1-2 weeks after negative nucleic acid conversion. However, there was no significant difference (χ 2 = 4.936, p = 0.552) in IgM concentration between time points tested (0-7 weeks) after negative nucleic acid conversion. The positive rates of IgM and IgG in asymptomatic patients (χ 2 = 84.660, p < 0.001) were significantly higher than those in the healthy control group (χ 2 = 9.201, p = 0.002) within 7 weeks of negative nucleic acid conversion. Conclusions:The IgG concentration in asymptomatic cases remained at a high level after nucleic acid turned negative. Nucleic acid detection combined with IgM and IgG antibody detection is an effective way to screen asymptomatic infections. How to cite this article: Li P, Shen G, Zhu Z, et al. Dynamic changes of IgM and IgG antibodies in asymptomatic patients as an effective way to detect SARS-CoV-2 infection.

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Section: Introductionmentioning

confidence: 99%

Dynamic changes of IgM and IgG antibodies in asymptomatic patients as an effective way to detect SARS‐CoV‐2 infection

Shen

Zhu

et al. 2021

Clinical Laboratory Analysis

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“…MCDA is as a novel isothermal amplification approach, first devised by Wang et al (2015) and is more sensitive than traditional PCR and other isothermal amplification methods. In the MCDA system, a suite of 10 specific primers with 10 binding sites for the target gene provide high specificity (Jiang et al, 2021;Luu et al, 2021). In our study, MCDA primers based on C. trachomatis ompA were successfully designed.…”

Section: Discussionmentioning

confidence: 99%

Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

Chen

Yuan²,

Zhou

et al. 2022

Front. Cell. Infect. Microbiol.

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Chlamydia trachomatis is the leading cause of bacterial sexually transmitted infection (STI) and remains a major public health challenge, especially in less-developed regions. Establishing a rapid, inexpensive, and easy-to-interpret point-of-care (POC) testing system for C. trachomatis could be critical for its treatment and limiting further transmission. Here, we devised a novel approach termed a multiple cross displacement amplification integrated with gold nanoparticle-based lateral flow biosensor (MCDA-AuNPs-LFB) for the highly specific, sensitive, user-friendly, and rapid identification of C. trachomatis in clinical samples. A suite of MCDA primers based on the C. trachomatis ompA gene from 14 serological variants (serovar A-K, L1, L2, and L3) were successfully designed and used to establish the assay. Optimal assay conditions were identified at 67°C, and the detection procedure, including nucleic acid preparation (approximately 5 min), MCDA amplification (30 min), and AuNPs-LFB visual readout (within 2 min), was completed within 40 min. The all-in cost for each test was approximately $5.5 USD. The limit of detection (LoD) was 10 copies/reaction, and no cross-reaction was observed with non-C. trachomatis microbes. A total of 135 suspected C. trachomatis-infection genital secretion samples were collected and simultaneously detected using real-time quantitative PCR (qPCR) in our assay. Compared with the qPCR technology, the MCDA-AuNPs-LFB sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 96.20%, 94.92%, and 100%, respectively. Hence, our MCDA-AuNP-LFB assay exhibited considerable potential for POC testing and could be used to identify C. trachomatis in clinical settings, particularly in low-income regions.

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“…MCDA (Multiple cross displacement amplification) is a method that amplifies DNA and makes use of ten primers, targeting ten distinct regions and a DNA polymerase with strand displacement activity ( Wang et al., 2015 ). MCDA is related to LAMP and when compared with it, enables faster results, despite a decrease in sensivity ( Luu et al., 2021 ). This method has been explored for diagnostic of SARS-CoV-2 in a consistent way, being mentioned in various works where it is combined with RT enzyme, despite the inexistence of a commercial assay based on the method ( Luu et al., 2021 ; Zhu et al., 2021 ).…”

Section: Nucleic Acids Amplificationmentioning

confidence: 99%

“…MCDA is related to LAMP and when compared with it, enables faster results, despite a decrease in sensivity ( Luu et al., 2021 ). This method has been explored for diagnostic of SARS-CoV-2 in a consistent way, being mentioned in various works where it is combined with RT enzyme, despite the inexistence of a commercial assay based on the method ( Luu et al., 2021 ; Zhu et al., 2021 ).…”

Section: Nucleic Acids Amplificationmentioning

confidence: 99%

SARS-CoV-2 Diagnostics Based on Nucleic Acids Amplification: From Fundamental Concepts to Applications and Beyond

Vindeirinho

Pinho

Azevedo

et al. 2022

Front. Cell. Infect. Microbiol.

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COVID-19 pandemic ignited the development of countless molecular methods for the diagnosis of SARS-CoV-2 based either on nucleic acid, or protein analysis, with the first establishing as the most used for routine diagnosis. The methods trusted for day to day analysis of nucleic acids rely on amplification, in order to enable specific SARS-CoV-2 RNA detection. This review aims to compile the state-of-the-art in the field of nucleic acid amplification tests (NAATs) used for SARS-CoV-2 detection, either at the clinic level, or at the Point-Of-Care (POC), thus focusing on isothermal and non-isothermal amplification-based diagnostics, while looking carefully at the concerning virology aspects, steps and instruments a test can involve. Following a theme contextualization in introduction, topics about fundamental knowledge on underlying virology aspects, collection and processing of clinical samples pave the way for a detailed assessment of the amplification and detection technologies. In order to address such themes, nucleic acid amplification methods, the different types of molecular reactions used for DNA detection, as well as the instruments requested for executing such routes of analysis are discussed in the subsequent sections. The benchmark of paradigmatic commercial tests further contributes toward discussion, building on technical aspects addressed in the previous sections and other additional information supplied in that part. The last lines are reserved for looking ahead to the future of NAATs and its importance in tackling this pandemic and other identical upcoming challenges.

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Development and comparison of novel multiple cross displacement amplification (MCDA) assays with other nucleic acid amplification methods for SARS-CoV-2 detection

Cited by 16 publications

References 17 publications

Dynamic changes of IgM and IgG antibodies in asymptomatic patients as an effective way to detect SARS‐CoV‐2 infection

Dynamic changes of IgM and IgG antibodies in asymptomatic patients as an effective way to detect SARS‐CoV‐2 infection

Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

SARS-CoV-2 Diagnostics Based on Nucleic Acids Amplification: From Fundamental Concepts to Applications and Beyond

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