Evaluation of GBV-C / HVG viremia in HIV-infected women

The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

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Validation of a Real-Time Quantitative Polymerase Chain Reaction Method for the Quantification of 3 Survivin Transcripts and Evaluation in Breast Cancer Tissues

Clinical Breast Cancer ◽

10.1016/j.clbc.2013.10.012 ◽

2014 ◽

Vol 14 (2) ◽

pp. 122-131 ◽

Cited By ~ 7

Author(s):

Anastasia Pavlidou ◽

Christos Kroupis ◽

Nikolaos Goutas ◽

Maria Dalamaga ◽

Kleanthi Dimas

Keyword(s):

Breast Cancer ◽

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Polymerase Chain Reaction Method ◽

Chain Reaction ◽

Reaction Method ◽

Polymerase Chain ◽

Cancer Tissues

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Midturbinate Swabs Are Comparable to Nasopharyngeal Swabs for Quantitative Detection of Respiratory Syncytial Virus in Infants

Journal of the Pediatric Infectious Diseases Society ◽

10.1093/jpids/piy115 ◽

2018 ◽

Vol 8 (6) ◽

pp. 554-558 ◽

Cited By ~ 2

Author(s):

Anne J Blaschke ◽

Matt McKevitt ◽

Krow Ampofo ◽

Tammi Lewis ◽

Hao Chai ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Respiratory Syncytial Virus ◽

Real Time ◽

Reverse Transcription ◽

Quantitative Polymerase Chain Reaction ◽

Quantitative Detection ◽

Chain Reaction ◽

Viral Loads ◽

Polymerase Chain ◽

Syncytial Virus

Abstract Nasopharyngeal (NP) swabs are generally used to detect respiratory syncytial virus (RSV) in infants. However, midturbinate (MT) swabs may provide comparable results. In this study, we enrolled hospitalized infants aged <24 months with RSV and collected NP and MT swabs. The resulting viral loads measured by real-time reverse-transcription quantitative polymerase chain reaction were similar. Most parents preferred MT swabs over NP swabs.

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Identification of Cervidae DNAin Feedstuff Using a Real-Time Polymerase Chain Reaction Method with the New Fluorescence Intercalating Dye EvaGreen

Journal of AOAC International ◽

10.1093/jaoac/92.1.175 ◽

2009 ◽

Vol 92 (1) ◽

pp. 175-180 ◽

Cited By ~ 5

Author(s):

Ying Chen ◽

Yajun Wu ◽

Jing Wang ◽

Baoliang Xu ◽

Zhengyu Zhong ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Polymerase Chain Reaction Method ◽

Chain Reaction ◽

Reaction Method ◽

Meat Meal ◽

Hair Samples ◽

Intercalating Dye ◽

Polymerase Chain

Abstract A real-time qualitative and quantitative polymerase chain reaction method (cer-194) using the fluorescence dye EvaGreen and aimed at the cytochrome b sequence was established for detection of cervidae DNA in feedstuff. Eight meat meal samples derived from deer, bovine, ovine, camel, pig, rabbit, fish, and chicken and 17 cervidae hair samples covering 2 subfamilies, 4 genera, and 7 species were tested to prove the specificity of the cer-194 system and its universality within the cervidae family. Detection limit of 0.1 deer meat in fish meal, blood powder, and feather powder matrixes was confirmed.

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TaqMan-based, real-time quantitative polymerase chain reaction method for RNA editing analysis

Analytical Biochemistry ◽

10.1016/j.ab.2009.04.011 ◽

2009 ◽

Vol 390 (2) ◽

pp. 173-180 ◽

Cited By ~ 5

Author(s):

Kevin Wong ◽

Rebecca Lyddon ◽

Stella Dracheva

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Rna Editing ◽

Quantitative Polymerase Chain Reaction ◽

Polymerase Chain Reaction Method ◽

Chain Reaction ◽

Reaction Method ◽

Polymerase Chain

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Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

Virology Journal ◽

10.1186/s12985-020-01467-y ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Yang Zhang ◽

Chunyang Dai ◽

Huiyan Wang ◽

Yong Gao ◽

Tuantuan Li ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Clinical Samples ◽

Chain Reaction ◽

Pcr Assay ◽

Qrt Pcr ◽

Highly Sensitive ◽

One Step ◽

Polymerase Chain

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.

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