Highly sensitive and fast detection of Phoma tracheiphila by polymerase chain reaction

1990 ◽  
Vol 32 (5) ◽  
Author(s):  
Franco Rollo ◽  
Roberto Salvi ◽  
Pietro Torchia
2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.


2004 ◽  
Vol 18 (16) ◽  
pp. 775-784 ◽  
Author(s):  
DIETER BRAUN

The Polymerase Chain Reaction (PCR) allows for highly sensitive and specific amplification of DNA. It is the backbone of many genetic experiments and tests. Recently, three labs independently uncovered a novel and simple way to perform a PCR reaction. Instead of repetitive heating and cooling, a temperature gradient across the reaction vessel drives thermal convection. By convection, the reaction liquid circulates between hot and cold regions of the chamber. The convection triggers DNA amplification as the DNA melts into two single strands in the hot region and replicates into twice the amount in the cold region. The amplification progresses exponentially as the convection moves on. We review the characteristics of the different approaches and show the benefits and prospects of the method.


2020 ◽  
Vol 10 (2) ◽  
pp. 90-92
Author(s):  
Rano Mal Piryani ◽  
Suneel Piryani ◽  
Shomeeta Piryani ◽  
Ganesh Dangal ◽  
Muzaherul Huq ◽  
...  

COVID-19 is mainly transmitted through droplet infection and spread very fast compared to SARS-CoV and MERS-CoV. For the countries, it is important to know at what stage the COVID-19 epidem­ic is? So, as to take appropriate steps to contain the epidemic. This will only be known by testing the suspects and contacts of confirmed cases. If there is poor testing, then most of the infected people may remain undetected, however they could spread the virus to hundreds of other people and potential contacts, which could not be known and quarantined in time continuing the spread. If there is quality assured, highly sensitive and specific testing along with adequate isolation and quarantine, then the spread will be limited. There are two types of tests available for COVID-19: the tests directly detecting the viral ribonucleic acid (RNA) collected in nasopharyngeal or throat swabs, and tests detecting antibodies from the blood sample. At this point in time, the polymerase chain reaction (PCR) tests are used for confirmation of the disease while antibodies tests may provide information regarding the prevalence of infection. World Health Organization advices the countries to increase the testing and get to know the level of epidemic and act accordingly for containment of infection.


Acta Tropica ◽  
2005 ◽  
Vol 94 (1) ◽  
pp. 55-59 ◽  
Author(s):  
Jene Greyce S. Oliveira ◽  
Fernanda O. Novais ◽  
Camila I. de Oliveira ◽  
Antonio C. da Cruz Junior ◽  
Léon Fábio Campos ◽  
...  

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