Combinations of PCR and isothermal amplification techniques are suitable for fast and sensitive detection of SARS-CoV-2 viral RNA

Varlamov, Dmitriy A; Blagodatskikh, Konstantin A.; Smirnova, Evgeniya V.; Kramarov, V. M.; Ignatov, Konstantin B.

doi:10.1101/2020.09.23.20200055

Cited by 3 publications

(6 citation statements)

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“…The RT-PCR test results obtained from nasopharyngeal, oropharyngeal or swap samples from lower respiratory tract such as sputum, tracheal aspirate, or bronchoalveolar lavage are considered variable and potentially unstable (10,12). RT-PCR is the primary method for the diagnosis of COVID-19, but it can cause false negativity (7,13,14). In a study in which Li et al (10) investigated the positivity of the RT-PCR test in patients diagnosed with COVID-19 clinically, they emphasized that there was a high rate of false negativity in the RT-PCR test.…”

Section: Discussionmentioning

confidence: 99%

BT Görüntüleme Bulgularına Göre Tanı Alan COVID-19 Pozitif ve Olası Hastaları Karşılaştırarak Klinik Karar Vermede BT'nin Kullanılabilirliğinin Araştırılması

Tekin

Bayraktar

Gür

et al. 2021

Düzce Tıp Fakültesi Dergisi

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Aim: In this study, the usability of thoracic computed tomography (CT) in clinical decision making was investigated by comparing laboratory results of patients with probable and definite coronavirus disease 2019 (COVID-19) diagnosis according to CT imaging features. Material and Methods: Within the scope of this single-center retrospective clinical study, data of possible and definite cases of COVID-19 were scanned from the hospital electronic database and patient files. Laboratory and CT imaging results of the patients were obtained. Patients were divided into two groups as positive and negative according to their CT imaging results, and compared. Results: Of the 995 patients included in the study, 57% (n=567) were male, and the mean age was 45.7±20.2 years. It was found that 65.1% (n=648) of the patients had positive CT. Realtime polymerase chain reaction (RT-PCR) test result was found positive in 22.2% (n=144) of the CT positive patients, and 32.0% (n=111) of the CT negative patients, and it was statistically significant (p<0.001). In the logistic regression analysis, it was determined that C-reactive protein (CRP), lymphocyte count, ferritin, procalcitonin, D-dimer, lactate and RT-PCR were statistically significant with CT positivity. Conclusion:In this study, COVID-19 positive and probable patients were compared according to thoracic CT findings and the usability of CT for clinical decision making was investigated. It has been determined that thorax CT can be used to initiate the treatment of COVID-19 in patients with negative RT-PCR test results but positive CT findings and high biochemical parameters such as CRP, D-dimer, ferritin and lactate.

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Section: Discussionmentioning

confidence: 99%

BT Görüntüleme Bulgularına Göre Tanı Alan COVID-19 Pozitif ve Olası Hastaları Karşılaştırarak Klinik Karar Vermede BT'nin Kullanılabilirliğinin Araştırılması

Tekin

Bayraktar

Gür

et al. 2021

Düzce Tıp Fakültesi Dergisi

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“…171 Several reports proved with the experimental evidence that a LOD as low as two copies can be achieved using a CRISPR-based platform. 32,65 The total testing time using the CRISPR-based platform varies based on the sensor platform utilized for the sensing purposes, ranging from 15 min to ∼1 h. 172−174 The CRISPR-based sensor can be further adapted to detect the infection caused by several The use of nanomaterials can advance the performance of the sensing technologies in virus identification. The integration of nanomaterials into a sensor device, e.g., quartz crystal microbalance, surface acoustic wave (SAW), or chemiresistors, led to many advancements in the field of sensing.…”

Section: Acs Nano Wwwacsnanoorgmentioning

confidence: 99%

“…63,64 Recently, a combination of PCR and isothermal nucleic acid amplification known as loop-mediated isothermal amplification (LAMP) techniques was explored. 65 The idea was to introduce the speed of the LAMP techniques and the accuracy of the PCR method in one single platform. It has been shown that combining LAMP and RT-PCR in one assay to develop RTq(PCR-LAMP) was approximately 100-fold more sensitive than conventional RT-LAMP.…”

Section: Nucleic Acid Amplification Test (Naat)mentioning

confidence: 99%

“…Using RT-q(PCR-LAMP) for COVID-19 diagnosis enables the detection of SARS-CoV-2 RNA of as few as five copies per reaction within a short time of 35 min for the amplification step (six cycles of PCR). 65 In another attempt, Shirato et al 66 explored a real-time RT-PCRbased assay for the ultrarapid detection of SARS-CoV-2 using the PCR1100 device. The whole amplification procedure takes less than 20 min and could achieve a sensitivity and specificity comparable to those of conventional real-time RT-PCR performed using thermocycler instruments.…”

Section: Nucleic Acid Amplification Test (Naat)mentioning

confidence: 99%

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Diagnostic Approaches For COVID-19: Lessons Learned and the Path Forward

Alafeef

Pan

2022

ACS Nano

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Coronavirus disease 2019 (COVID-19) is a transmitted respiratory disease caused by the infection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although humankind has experienced several outbreaks of infectious diseases, the COVID-19 pandemic has the highest rate of infection and has had high levels of social and economic repercussions. The current COVID-19 pandemic has highlighted the limitations of existing virological tests, which have failed to be adopted at a rate to properly slow the rapid spread of SARS-CoV-2. Pandemic preparedness has developed as a focus of many governments around the world in the event of a future outbreak. Despite the largely widespread availability of vaccines, the importance of testing has not diminished to monitor the evolution of the virus and the resulting stages of the pandemic. Therefore, developing diagnostic technology that serves as a line of defense has become imperative. In particular, that test should satisfy three criteria to be widely adopted: simplicity, economic feasibility, and accessibility. At the heart of it all, it must enable early diagnosis in the course of infection to reduce spread. However, diagnostic manufacturers need guidance on the optimal characteristics of a virological test to ensure pandemic preparedness and to aid in the effective treatment of viral infections. Nanomaterials are a decisive element in developing COVID-19 diagnostic kits as well as a key contributor to enhance the performance of existing tests. Our objective is to develop a profile of the criteria that should be available in a platform as the target product. In this work, virus detection tests were evaluated from the perspective of the COVID-19 pandemic, and then we generalized the requirements to develop a target product profile for a platform for virus detection.

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“…Loop-mediated isothermal amplification (LAMP): In an LAMP experiment, four to six primers are specially designed to identify six to eight regions of the target sequence. 56,57 The amplification begins with an inner primer invasion, then a strand-displacing polymerase elongates the primer and separates the DNA duplex. Subsequently, the first product is displaced by another strand, which is extended by an outer primer.…”

Section: Introductionmentioning

confidence: 99%

Applications of laboratory findings in the prevention, diagnosis, treatment, and monitoring of COVID-19

Meng

Guo

Zhou

et al. 2021

Sig Transduct Target Ther

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The worldwide pandemic of coronavirus disease 2019 (COVID-19) presents us with a serious public health crisis. To combat the virus and slow its spread, wider testing is essential. There is a need for more sensitive, specific, and convenient detection methods of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Advanced detection can greatly improve the ability and accuracy of the clinical diagnosis of COVID-19, which is conducive to the early suitable treatment and supports precise prophylaxis. In this article, we combine and present the latest laboratory diagnostic technologies and methods for SARS-CoV-2 to identify the technical characteristics, considerations, biosafety requirements, common problems with testing and interpretation of results, and coping strategies of commonly used testing methods. We highlight the gaps in current diagnostic capacity and propose potential solutions to provide cutting-edge technical support to achieve a more precise diagnosis, treatment, and prevention of COVID-19 and to overcome the difficulties with the normalization of epidemic prevention and control.

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Combinations of PCR and isothermal amplification techniques are suitable for fast and sensitive detection of SARS-CoV-2 viral RNA

Cited by 3 publications

References 25 publications

BT Görüntüleme Bulgularına Göre Tanı Alan COVID-19 Pozitif ve Olası Hastaları Karşılaştırarak Klinik Karar Vermede BT'nin Kullanılabilirliğinin Araştırılması

BT Görüntüleme Bulgularına Göre Tanı Alan COVID-19 Pozitif ve Olası Hastaları Karşılaştırarak Klinik Karar Vermede BT'nin Kullanılabilirliğinin Araştırılması

Diagnostic Approaches For COVID-19: Lessons Learned and the Path Forward

Applications of laboratory findings in the prevention, diagnosis, treatment, and monitoring of COVID-19

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