How to Avoid False-Negative and False-Positive COVID-19 PCR Testing

Fevraleva, I S; Глинщикова, О А; Makarik, Tatiana; Sudarikov, Andrey

doi:10.3390/ijtm2020018

Cited by 4 publications

(4 citation statements)

References 9 publications

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“…Another PCR assay using the same genes was effective in identifying 95% of P. aeruginosa isolates from Dorper sheep milk 21 . Multiplex PCR can also be developed to allow for more in-depth diagnostics, as multiple bands and single bands can be interpreted differently, minimizing false positives and false negatives 22 . This study considers the possibility of a duplex PCR detection method using oprL coupled with algD or nfxB genes.…”

Section: Discussionmentioning

confidence: 99%

Identification of commensal Pseudomonas aeruginosa isolates using duplex PCR targeting the oprL and algD genes

et al. 2022

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Background: Pseudomonas aeruginosa (P. aeruginosa) is a ubiquitous bacterium that can be found in most moisture places, such as soil, water, food, plants, and animals, including humans. Due to genetic flexibility among strains, there is no standard molecular identification for P. aeruginosa from different sources. In this study, monoplex and duplex PCR targeting oprL, algD and nfxB were assessed for the identification of commensal P. aeruginosa. Methods: Twenty-nine commensal Pseudomonas isolates, including 16 P. aeruginosa isolates and 13 P. aeruginosa-like isolates, were used in the study. First, monoplex PCR targeting oprL, algD and nfxB using published primers was carried out to test their ability to detect commensal Pseudomonas isolates. Then, two new primer pairs targeting oprL were designed (oprL-pp1 and oprL-pp2) and used for oprL-algD duplex PCR to check for the improvement of commensal P. aeruginosa detection. Result and conclusion: AlgD or nfxB monoplex PCR had the same sensitivity of 93.75% and specificity of 100%, while oprL PCR using published primers was more sensitive (100%) but less specific (0%). Duplex PCR yielded high sensitivity and specificity in detecting P. aeruginosa. Both oprL-pp1/algD and oprL-pp2/algD duplex-PCR had 93.75% sensitivity (15/16 P. aeruginosa isolates) and 100% specificity (0/13 P. aeruginosa-like isolates). In addition, oprL-pp2 primers were more specific than oprL-pp1 primers, with only 2 of 13 P. aeruginosa-like isolates detected, while oprL-pp1 primers detected all P. aeruginosa-like isolates. Compared to the monoplex PCR that only targeted the oprL gene, the duplex PCR utilizing oprL-pp2 and algD primers identified 15/16 P. aeruginosa isolates (93.75% specificity). Additionally, the duplex PCR used in this study was negative for non-Pseudomonas species, including E. coli, V. cholera, V. parahaemolyticus, and S. aureus. In conclusion, our duplex PCR targeting oprL and algD could be a valuable tool for commensal P. aeruginosa screening.

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Section: Discussionmentioning

confidence: 99%

Identification of commensal Pseudomonas aeruginosa isolates using duplex PCR targeting the oprL and algD genes

et al. 2022

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“…Finally, the process completes with the quantitative analysis of the viral load. However, it is prone to resulting in false positives or false negatives [ 32 , 33 , 34 ]. To address these issues, the research by Xie and colleagues has led to the creation of a novel three-stage microfluidic system that integrates both reverse transcription and pre-amplification, significantly increasing the detection capacity for low-viral-load samples and solving the false-negative issue.…”

Section: The Application Of Pcr Technology In Microfluidic Platformsmentioning

confidence: 99%

Advances in Nucleic Acid Assays for Infectious Disease: The Role of Microfluidic Technology

Wang,

Chen,

Yang

et al. 2024

Molecules

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Within the fields of infectious disease diagnostics, microfluidic-based integrated technology systems have become a vital technology in enhancing the rapidity, accuracy, and portability of pathogen detection. These systems synergize microfluidic techniques with advanced molecular biology methods, including reverse transcription polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), and clustered regularly interspaced short palindromic repeats (CRISPR), have been successfully used to identify a diverse array of pathogens, including COVID-19, Ebola, Zika, and dengue fever. This review outlines the advances in pathogen detection, attributing them to the integration of microfluidic technology with traditional molecular biology methods and smartphone- and paper-based diagnostic assays. The cutting-edge diagnostic technologies are of critical importance for disease prevention and epidemic surveillance. Looking ahead, research is expected to focus on increasing detection sensitivity, streamlining testing processes, reducing costs, and enhancing the capability for remote data sharing. These improvements aim to achieve broader coverage and quicker response mechanisms, thereby constructing a more robust defense for global public health security.

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“…Therefore, meticulous attention to primer design is crucial for addressing these challenges. 11,12 Scheme 1 (A) Schematic diagram of the common and differential gene screening on the Wild strains Y3 and A19 worldwide from the brucella. P1, which are complementary to T1, is used for screening positive samples of brucella (the black elliptic).…”

Section: Introductionmentioning

confidence: 99%

Gated nanoprobe utilizing metal–organic frameworks for identifying and distinguishing between the wild strains and the vaccine strains of brucella

Li,

Ren,

Wang

et al. 2024

Analyst

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How to Avoid False-Negative and False-Positive COVID-19 PCR Testing

Cited by 4 publications

References 9 publications

Identification of commensal Pseudomonas aeruginosa isolates using duplex PCR targeting the oprL and algD genes

Identification of commensal Pseudomonas aeruginosa isolates using duplex PCR targeting the oprL and algD genes

Advances in Nucleic Acid Assays for Infectious Disease: The Role of Microfluidic Technology

Gated nanoprobe utilizing metal–organic frameworks for identifying and distinguishing between the wild strains and the vaccine strains of brucella

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