scholarly journals maxRatio improves the detection of samples with abnormal amplification profiles on QIAgen’s artus HIV-1 qPCR assay

F1000Research ◽  
2021 ◽  
Vol 9 ◽  
pp. 1030
Author(s):  
Luigi Marongiu ◽  
Eric B. Shain ◽  
Marianna Martinelli ◽  
Matteo Pagliari ◽  
Heike Allgayer
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Predictive Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Alternative Approach ◽  
External Standard ◽  
Polymerase Chain ◽  
Anti Hiv ◽  
Hiv 1

Background: Accurate viral load (VL) determination is paramount to determine the efficacy of anti-HIV-1 therapy. The conventional method used, fit-point (FP), assumes an equal efficiency in the polymerase chain reaction (PCR) among samples that might not hold for low-input templates. An alternative approach, maxRatio, was introduced to compensate for inhibition in PCR. Methods: Herein, we assessed whether maxRatio could improve VL quantification using 2,544 QIAgen artus HI virus-1 RT-PCR reactions. The assay’s standard dilutions were used to build external standard curves with either FP or maxRatio that re-calculated the VLs. Results: FP and maxRatio were highly comparable (Pearson’s ρ=0.994, Cohen’s  κ=0.885), and the combination of the two methods identified samples (n=41) with aberrant amplification profiles. Conclusions: The combination of maxRatio and FP could improve the predictive value of the assay.

scholarly journals maxRatio improves the detection of samples with abnormal amplification profiles on QIAgen’s artus HIV-1 qPCR assay

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 1030
Author(s):  
Luigi Marongiu ◽  
Eric B. Shain ◽  
Marianna Martinelli ◽  
Matteo Pagliari ◽  
Heike Allgayer
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Predictive Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Alternative Approach ◽  
External Standard ◽  
Polymerase Chain ◽  
Anti Hiv ◽  
Hiv 1

Background: Accurate viral load (VL) determination is paramount to determine the efficacy of anti-HIV-1 therapy. The conventional method used, fit-point (FP), assumes an equal efficiency in the polymerase chain reaction (PCR) among samples that might not hold for low-input templates. An alternative approach, maxRatio, was introduced to compensate for inhibition in PCR. Methods: Herein, we assessed whether maxRatio could improve VL quantification using 2,544 QIAgen artus HI virus-1 RT-PCR reactions. The assay’s standard dilutions were used to build external standard curves with either FP or maxRatio that re-calculated the VLs. Results: FP and maxRatio were highly comparable (Pearson’s ρ=0.994, Cohen’s  κ=0.885), and the combination of the two methods identified samples (n=41) with aberrant amplification profiles. Conclusions: The combination of maxRatio and FP could improve the predictive value of the assay.

scholarly journals maxRatio improves the detection of samples with abnormal amplification profiles on QIAgen’s artus HIV-1 qPCR assay

F1000Research ◽  
2021 ◽  
Vol 9 ◽  
pp. 1030
Author(s):  
Luigi Marongiu ◽  
Eric B. Shain ◽  
Marianna Martinelli ◽  
Matteo Pagliari ◽  
Heike Allgayer
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Predictive Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Alternative Approach ◽  
External Standard ◽  
Polymerase Chain ◽  
Anti Hiv ◽  
Hiv 1

Background: Accurate viral load (VL) determination is paramount to determine the efficacy of anti-HIV-1 therapy. The conventional method used, fit-point (FP), assumes an equal efficiency in the polymerase chain reaction (PCR) among samples that might not hold for low-input templates. An alternative approach, maxRatio, was introduced to compensate for inhibition in PCR. Methods: Herein, we assessed whether maxRatio could improve VL quantification using 2,544 QIAgen artus HI virus-1 RT-PCR reactions. The assay’s standard dilutions were used to build external standard curves with either FP or maxRatio that re-calculated the VLs. Results: FP and maxRatio were highly comparable (Pearson’s ρ=0.994, Cohen’s  κ=0.885), and the combination of the two methods identified samples (n=41) with aberrant amplification profiles. Conclusions: The combination of maxRatio and FP could improve the predictive value of the assay.

scholarly journals A meta-analysis of accuracy and sensitivity of chest CT and RT-PCR in COVID-19 diagnosis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Ct Scan ◽  
Predictive Value ◽  
Meta Analysis ◽  
Chest Ct ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chest Ct Scan

AbstractNowadays there is an ongoing acute respiratory outbreak caused by the novel highly contagious coronavirus (COVID-19). The diagnostic protocol is based on quantitative reverse-transcription polymerase chain reaction (RT-PCR) and chests CT scan, with uncertain accuracy. This meta-analysis study determines the diagnostic value of an initial chest CT scan in patients with COVID-19 infection in comparison with RT-PCR. Three main databases; PubMed (MEDLINE), Scopus, and EMBASE were systematically searched for all published literature from January 1st, 2019, to the 21st May 2020 with the keywords "COVID19 virus", "2019 novel coronavirus", "Wuhan coronavirus", "2019-nCoV", "X-Ray Computed Tomography", "Polymerase Chain Reaction", "Reverse Transcriptase PCR", and "PCR Reverse Transcriptase". All relevant case-series, cross-sectional, and cohort studies were selected. Data extraction and analysis were performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5. Among 1022 articles, 60 studies were eligible for totalizing 5744 patients. The overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan compared to RT-PCR were 87% (95% CI 85–90%), 46% (95% CI 29–63%), 69% (95% CI 56–72%), and 89% (95% CI 82–96%), respectively. It is important to rely on the repeated RT-PCR three times to give 99% accuracy, especially in negative samples. Regarding the overall diagnostic sensitivity of 87% for chest CT, the RT-PCR testing is essential and should be repeated to escape misdiagnosis.

scholarly journals Performance of the Innova SARS-CoV-2 antigen rapid lateral flow test in the Liverpool asymptomatic testing pilot: population based cohort study

BMJ ◽  
2021 ◽  
pp. n1637 ◽  
Author(s):  
Marta García-Fiñana ◽  
David M Hughes ◽  
Christopher P Cheyne ◽  
Girvan Burnside ◽  
Mark Stockbridge ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Predictive Value ◽  
High Viral Load ◽  
Test Results ◽  
Chain Reaction ◽  
Viral Loads ◽  
Flow Test ◽  
Polymerase Chain ◽  
Lateral Flow Test

Abstract Objective To assess the performance of the SARS-CoV-2 antigen rapid lateral flow test (LFT) versus polymerase chain reaction testing in the asymptomatic general population attending testing centres. Design Observational cohort study. Setting Community LFT pilot at covid-19 testing sites in Liverpool, UK. Participants 5869 asymptomatic adults (≥18 years) voluntarily attending one of 48 testing sites during 6-29 November 2020. Interventions Participants were tested using both an Innova LFT and a quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) test based on supervised self-administered swabbing at testing sites. Main outcome measures Sensitivity, specificity, and predictive values of LFT compared with RT-qPCR in an epidemic steady state of covid-19 among adults with no classic symptoms of the disease. Results Of 5869 test results, 22 (0.4%) LFT results and 343 (5.8%) RT-qPCR results were void (that is, when the control line fails to appear within 30 minutes). Excluding the void results, the LFT versus RT-qPCR showed a sensitivity of 40.0% (95% confidence interval 28.5% to 52.4%; 28/70), specificity of 99.9% (99.8% to 99.99%; 5431/5434), positive predictive value of 90.3% (74.2% to 98.0%; 28/31), and negative predictive value of 99.2% (99.0% to 99.4%; 5431/5473). When the void samples were assumed to be negative, a sensitivity was observed for LFT of 37.8% (26.8% to 49.9%; 28/74), specificity of 99.6% (99.4% to 99.8%; 5431/5452), positive predictive value of 84.8% (68.1% to 94.9%; 28/33), and negative predictive value of 93.4% (92.7% to 94.0%; 5431/5814). The sensitivity in participants with an RT-qPCR cycle threshold (Ct) of <18.3 (approximate viral loads >10 6 RNA copies/mL) was 90.9% (58.7% to 99.8%; 10/11), a Ct of <24.4 (>10 4 RNA copies/mL) was 69.4% (51.9% to 83.7%; 25/36), and a Ct of >24.4 (<10 4 RNA copies/mL) was 9.7% (1.9% to 23.7%; 3/34). LFT is likely to detect at least three fifths and at most 998 in every 1000 people with a positive RT-qPCR test result with high viral load. Conclusions The Innova LFT can be useful for identifying infections among adults who report no symptoms of covid-19, particularly those with high viral load who are more likely to infect others. The number of asymptomatic adults with lower Ct (indicating higher viral load) missed by LFT, although small, should be considered when using single LFT in high consequence settings. Clear and accurate communication with the public about how to interpret test results is important, given the chance of missing some cases, even at high viral loads. Further research is needed to understand how infectiousness is reflected in the viral antigen shedding detected by LFT versus the viral loads approximated by RT-qPCR.

Prognostic Value of Circulating Melanoma Cells Detected by Reverse Transcriptase–Polymerase Chain Reaction

2003 ◽  
Vol 21 (5) ◽  
pp. 767-773 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Paolo A. Ascierto ◽  
Francesco Perrone ◽  
Sabrina M.R. Satriano ◽  
Alessandro Ottaiano ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Subgroup Analysis ◽  
Peripheral Blood ◽  
Predictive Value ◽  
Prognostic Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Stage Of Disease ◽  
Polymerase Chain

Purpose: Factors that are predictive of prognosis in patients who are diagnosed with malignant melanoma (MM) are widely awaited. Detection of circulating melanoma cells (CMCs) by reverse transcriptase-polymerase chain reaction (RT-PCR) has recently been postulated as a possible negative prognostic factor. Two main questions were addressed: first, whether the presence of CMCs, defined as the patient being positive for any of the three markers, had a prognostic role; and second, what the predictive value of each individual marker was. Patients and Methods: A consecutive series of 200 melanoma patients observed between January 1997 and December 1997, with stage of disease ranging from I to IV, was analyzed by semiquantitative RT-PCR. Tyrosinase, p97, and MelanA/MART1 were used as markers to CMCs on baseline peripheral blood samples. Progression-free survival (PFS) was used as a unique end point and was described by the product limit method. Multivariable analysis was applied to verify whether the auspicated prognostic value of these markers was independent of the stage of disease, and a subgroup analysis was performed that excluded patients with stage IV disease. Results: Overall, 32% (64 of 200) of patients progressed, and a median PFS of 52 months in the whole series was observed. The presence of CMCs and the markers individually or combined was predictive of prognosis in the univariate analysis but did not provide additional prognostic information to the stage of disease in multivariable models. In the subgroup analysis of stage (ie, I–III subgroup), similar results were observed. Conclusion: Detection of CMCs in peripheral blood samples at the time of MM diagnosis by semiquantitative RT-PCR does not add any significant predictive value to the stage of disease. Thus, this approach should not be used in clinical practice, and further studies are required to determine its usefulness.

scholarly journals The utility of real-time polymerase chain reaction in detecting Coccidioides immitis among clinical specimens in the Central California San Joaquin Valley

2018 ◽  
Vol 57 (6) ◽  
pp. 688-693 ◽  
Author(s):  
Dominic Dizon ◽  
Marilyn Mitchell ◽  
Bernadette Dizon ◽  
Robert Libke ◽  
Michael W Peterson
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Pleural Fluid ◽  
Clinical Setting ◽  
Predictive Value ◽  
Coccidioides Immitis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Acute Pneumonia ◽  
Polymerase Chain

AbstractCoccidioidomycosis, the fungal infection caused by dimorphic Coccidioides species, is typically diagnosed by histopathologic identification of spherules, by culture, or by serology. These tests are reliable but time-intensive, delaying diagnosis and treatment. Rapid real-time polymerase chain reaction (RT-PCR) can be performed and was validated to identify Coccidioides immitis using an in-house developed assay for the Becton Dickinson molecular instrument (BD MAXTM). These studies were performed using patient samples that had been shown to be positive on previously set up fungal cultures. To evaluate this new RT-PCR test in the clinical setting, we conducted a retrospective chart review of patients (N = 1160) who underwent Coccidioides PCR (Cocci PCR) on clinical samples between March 1, 2014, and Dec 31, 2016. We abstracted clinical, microbiologic, serologic, radiographic, treatment, and follow-up data. Specimens of cerebrospinal fluid (CSF), bronchioalveolar lavage fluid (BAL), lung tissue biopsy (LTB), sputum, and pleural fluid were evaluated to determine sensitivity and specificity. Of the 113 specimens that tested positive for Cocci PCR, all had clinical disease defined by traditional clinical criteria, yielding 100% specificity. Overall sensitivity was 74% versus 46% for fungal culture and was available in 4 hours rather than 1–2 weeks. Sensitivities varied by source material and clinical setting. CSF had a sensitivity of 59%, BAL for acute pneumonia 91%, sputum for acute pneumonia 94%, pleural fluid 86%, but LTB for lung nodules only 44%. Overall positive predictive value (PPV) was 100%, while negative predictive value (NPV) was 96%, but again this varied by specimen and clinical setting. Our experience with clinical testing of >1160 specimens over 2–3 years shows we can utilize this technology to improve our ability to diagnose disease but that the sensitivity varies by specimen source and clinical setting.

scholarly journals Is the Portable Chest Radiographic More Reliable to Reveal Covid19 in Highly Suspicion Patient before Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) Test?

2021 ◽  
Vol 6 (03) ◽  
pp. 228-236
Author(s):  
Bushra A. A. Albazi ◽  
Dr Noof. Albaz ◽  
Dr Nayef. Alqahtani ◽  
Dr. Angham Salih ◽  
Dr Rafat Mohtasab
Keyword(s):  
Polymerase Chain Reaction ◽  
Retrospective Study ◽  
Predictive Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
X Ray ◽  
Polymerase Chain ◽  
Rate Type ◽  
Test Result ◽  
Pcr Test

A large number of patients with coronavirus disease 2019 (COVID-19) present at hospitals. There are a limited number of isolation rooms open, and patients must often wait a long time to get a reverse transcription-polymerase chain reaction (RT-PCR) test done. This necessitates the introduction of effective triage plans. A patient with suspicions is referred to an emergency room (ED) depending on their medical record for a simple physical assessment, blood test findings, and chest imaging.A retrospective study design was conduct at Prince Sultan Medical Military City (PSMMC). Ethical approval was obtained from the institutional board to wave the consent forms since it is a retrospective study. Only the primary investigator has had the data access to the patients’ medical records. The collected patient records were under specific categories, including symptoms score starts from 5 and above, RT-PCR test result done after CXRP imaging, the patient admitted to the emergency department (ED). Excluding all CXRP done after RT-PCR TEST, positive Covid 19 admitted to the intensive care unit (ICU), pediatric patients, and patients with score symptoms were less than five. Two experienced radiologists reviewed the images blindly, and the inter-observer reliability of observations noted by the radiologists was calculated. As for the relationship between the x-ray reading and the RT-PCR test result, our results showed a high correlation between the variables (chi-square χ² = 12.44, with df =1, and p<0.001). The sensitivity of x-ray diagnosing covid19 was 65.52 %, while the specificity was 54.51 %, and the accuracy of radiologists reading was 58.17 %. Furthermore, the positive predictive value (PPV) was 41.76 %, and the negative predictive value (NPV) was 76.05%. Finally, the false positive rate (type-i error (alpha) was 45.49%, and the false-negative rate (type-ii error (beta) was 34.48% Our research findings show that CXRP imaging can detect COVID-19 infection in symptomatic patients and can be a valuable addition to RT-PCR testing. In an inpatient ED environment where availability of test kits, laboratory equipment, and laboratory personnel is compromised and risks delaying patient treatment and hospital workflow, serial CXRP could theoretically be used as an adjunct diagnostic function and monitoring in patients suspected of having COVID-19.

scholarly journals Performance comparison of two malaria rapid diagnostic test with real time polymerase chain reaction and gold standard of microscopy detection method

2020 ◽  
Author(s):  
Puspa Wardhani ◽  
Trieva Verawaty Butarbutar ◽  
Christophorus Oetama Adiatmaja ◽  
Amarensi Milka Betaubun ◽  
Nur Hamidah ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Diagnostic Test ◽  
Real Time Pcr ◽  
Predictive Value ◽  
Gold Standard ◽  
Detection Method ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Background: The diagnostic test for malaria is mostly based on Rapid Diagnostic Test (RDT) and detection by microscopy. Polymerase Chain Reaction (PCR) is also a sensitive detection method that can be considered as a diagnostic tool. The outcome of malaria microscopy detection depends on the examiner's ability and experience. Some RDT has been distributed in Indonesia, which needs to be evaluated for their results. Objective: This study aimed to compare the performance of RightSign RDT and ScreenPlus RDT for detection of Plasmodium in human blood. We used specific real-time polymerase chain reaction abTESTMMalaria qPCRII) and gold standard of microscopy detection method to measure diagnostic efficiency. Methods: Blood specimens were evaluated using RightSign RDT, ScreenPlus RDT, Microscopy detection, and RT-PCR as the protocol described. The differences on specificity (Sp), sensitivity (Sn), positive predictive value (PPV), and negative predictive value (NPV) were analyzed using McNemar and Kruskal Wallis analysis. Results: A total of 105 subjects were recruited. Based on microscopy test, RightSign RDT had sensitivity, Specificity, PPV, NPV, 100%, 98%, 98.2%, 100%, respectively. ScreenPlus showed 100% sensitivity, 98% specificity, 98.2% PPV, 100% NPV. The sensitivity of both RDTs became lower (75%) and the specificity higher (100 %) when using real-time PCR. Both RDTs showed a 100% agreement. RT-PCR detected higher mix infection when compared to microscopy and RDTs. Conclusion: RightSign and ScreenPlus RDT have excellent performance when using microscopy detection as a gold standard. Real-time PCR method can be considered as a confirmation tool for malaria diagnosis.

scholarly journals The exact place of PCR and chest CT in screening, staging, and diagnosis of Covid-19: A meta-analysis

2020 ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Ct Scan ◽  
Predictive Value ◽  
Meta Analysis ◽  
Chest Ct ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chest Ct Scan

Abstract Introduction: Nowadays there is an ongoing acute respiratory outbreak causing by the novel highly contagious coronavirus (nCoV). There are two diagnostic protocol based on chest CT scan and quantitative reverse-transcription polymerase chain reaction (RT-PCR) which their diagnostic accuracy is under the debate. We designed this meta-analysis study to determine the diagnostic value of initial chest CT scan in patients with nCoV infection in comparison with RT- PCR.Search strategy and statistical analysis: Three main databases the PubMed (MEDLINE), Scopus, and EMBASE was systematically searched for all published literatures from January 1st, 2019, to the 27th march 2020 with key grouping of “COVID19 virus”, “2019 novel coronavirus”, “Wuhan coronavirus”, “2019-nCoV”, “X-Ray Computed Tomography”, “Polymerase Chain Reaction”, “Reverse Transcriptase PCR”, and “PCR Reverse Transcriptase”. All relevant case- series, cross-sectional, and cohort studies were selected. Data extraction was done in Excel 2007 (Microsoft Corporation, Redmond, CA) and their analysis was performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5.Result: From first recruited 668 articles we end up to the final 47 studies, which comprised a total sample size of 4238 patients. In compare to RT-PCR, the overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan were 86% (95% CI: 83% -90%), 43 % (95% CI: 26% -60%), 67% (95% CI: 57% -78%), and 84% (95% CI: 74% -95%) respectively. However the RT-PCR should be repeated for three times in order to give the 99% accuracy especially in negative samples.Conclusion: According to the acceptable sensitivity of chest CT scan, it can be employed complement to RT-PCR to diagnosis patients who are clinically suspicious for nCoV.

AN EPIDEMIOLOGY-CLINICAL CORRELATION OF VIRAL LOAD AS MEASURED BY CYCLE THRESHOLD VALUE BY RT-PCR IN COVID-1

2021 ◽  
pp. 1-2
Author(s):  
Atrikumar P. Patel ◽  
Palak Shah ◽  
Pavan Acharya ◽  
Monila N. Patel
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Severe Acute Respiratory Syndrome ◽  
Threshold Value ◽  
Clinical Correlation ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Oropharyngeal Swab ◽  
Polymerase Chain ◽  
Novel Coronavirus

The 2019 novel coronavirus [severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)] was rst documented in December 2019 in Wuhan, China, and spread across the globe resulting in [1]. signicant global morbidity and mortality Diagnosis of COVID-19 is mainly done by nasopharyngeal and oropharyngeal swab RT-PCR (Reverse transcriptase - polymerase chain reaction). Real time RT-PCR is of great interest today for detection of SARS- CoV-2 due to its benets as a specic assay.

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