Duplex Real-Time Reverse Transcriptase PCR Assays for Rapid Detection and Identification of Pandemic (H1N1) 2009 and Seasonal Influenza A/H1, A/H3, and B Viruses

Chidlow, Glenys; Harnett, Gerald B.; Williams, Simon H.; Levy, Avram; Speers, David; Smith, David W.

doi:10.1128/jcm.01435-09

Cited by 64 publications

(63 citation statements)

References 10 publications

(7 reference statements)

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“…[13][14] and Discussion sections (p.14) concerning these results to clarify this point. Figures 2 and 3 also include 95% confidence intervals to minimise the risk of over-interpretation.…”

Section: Responsementioning

confidence: 88%

“…All children 6-59 months of age presenting to PMH with a history of fever (by parental report) or with a measured temperature of greater than 37.5°C at [14,15]. Picornaviruses were detected using nested PCR [16] targeting the 5'UTR of the picornavirus genome with sequencing used to assist with identification of rhinoviruses and enteroviruses.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Viral Etiology and the Impact of Codetection in Young Children Presenting With Influenza-Like Illness

Lim

Wake²,

Levy³

et al. 2016

J Ped Infect Dis

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Citation for published version (APA):Lim, F. J., Wake, Z. V., Levy, A., Tempone, S., Moore, H. C., Richmond, P. C., ... Blyth, C. C. (2016). Viral etiology and the impact of co-detection in young children presenting with influenza-like illness. Journal of the Pediatric Infectious Disease Society, 6(3), 260-266. [piw042]. https://doi.org/10.1093/jpids/piw042General rights Copyright and moral rights for the publications made accessible in Discovery Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.• Users may download and print one copy of any publication from Discovery Research Portal for the purpose of private study or research.• You may not further distribute the material or use it for any profit-making activity or commercial gain.• You may freely distribute the URL identifying the publication in the public portal. ResponseAs the reviewer has suggested, the differences in the presented results in the Abstract were due to rounding. To minimise confusion, we have amended the abstract to report results up to 1 decimal place as appropriate (pp.5-6).The calculation of the predicted probabilities from the logistic regression model needs to be clearly described in the methods section. Whether the predicted probability for flu + RSV is significantly higher than others need to be clarified as well. ResponseWe have clarified this in the abstract of the revised manuscript (pp.5-6). Introduction Suggest reconcile description of study settings with information from methods, as apparently not all recruitments were done at the emergency department ResponseTo simplify analyses and minimise confusion regarding the study setting, we have chosen to exclude children enrolled from general practises in the revised manuscript. Only 131 were recruited from general practice in 2008-2009 before this arm of the study was stopped. All children included in the analyses were enrolled while they were transiting through the emergency department. A portion of these children would subsequently have been admitted as inpatients. We have included additional information on the timing and location of enrolment in the Introduction (p.7) with further information included in the Methods section (p.8). The number of patients excluded from general practice are included in the Results section (p.12). We acknowledge the confusion in the distribution of enrolment settings and consequently have opted to exclude data from all children enrolled at general practises (see response to Introduction comments above). All children presented to the emergency department of a single hospital and were either admitted or discharged home. Hospital admission was a key outcome of interest. Methods Settings and participants How was influenza vaccination data collected? What proportion of self-report was verified? What proportion of children were fully vaccinated?Response Vaccination data were collected through parental...

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“…[13][14] and Discussion sections (p.14) concerning these results to clarify this point. Figures 2 and 3 also include 95% confidence intervals to minimise the risk of over-interpretation.…”

Section: Responsementioning

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Viral Etiology and the Impact of Codetection in Young Children Presenting With Influenza-Like Illness

Lim

Wake²,

Levy³

et al. 2016

J Ped Infect Dis

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“…Methods currently used for detection include serological testing and immunological testing bioassays, polymerase chain reaction (PCR) and fluorescent quantitative real-time PCR (3,4). The mandated blood screenings in China are serological tests for HBV surface antigens, antibodies against T. pallidum, HCV, and HIV-1 (5).…”

Section: Introductionmentioning

confidence: 99%

Development of a Multiplex Real-Time PCR Assay for the Detection of <i>Treponema pallidum</i>, HCV, HIV-1, and HBV

et al. 2015

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SUMMARY: Treponema pallidum, hepatitis C virus (HCV), human immunodeficiency virus (HIV)-1, and hepatitis B virus (HBV) are major causes of sexually transmitted diseases passed through blood contact. The development of a sensitive and efficient method for detection is critical for early diagnosis and for large-scale screening of blood specimens in China. This study aims to establish an assay to detect these pathogens in clinical serum specimens. We established a TaqMan-locked nucleic acid (LNA) realtime polymerase chain reaction (PCR) assay for rapid, sensitive, specific, quantitative, and simultaneous detection and identification. The copy numbers of standards of these 4 pathogens were quantified. Standard curves were generated by determining the mean cycle threshold values versus 10-fold serial dilutions of standards over a range of 10 6 to 10 1 copies/mL, with the lowest detection limit of the assay being 10 1 copies/mL. The assay was applied to 328 clinical specimens and compared with enzyme-linked immunosorbent assay (ELISA) and commercial nucleic acid testing (NAT) methods. The assay identified 39 T. pallidum-, 96 HCV-, 13 HIV-1-, 123 HBV-, 5 HBV/HCV-, 1 T. pallidum/HBV-, 1 HIV-1/HCV-, and 1 HIV-1/T. pallidum-positive specimens. The high sensitivity of the assay confers strong potential for its use as a highly reliable, cost-effective, and useful molecular diagnostic tool for largescale screening of clinical specimens. This assay will assist in the study of the pathogenesis and epidemiology of sexually transmitted blood diseases.

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“…Similarly, culture has increased sensitivity over both RIDTs and DFAs but requires skilled technologists and specialized laboratory settings and has a long turnaround time (2 to 14 days) (17). NAAT are highly sensitive and are gradually replacing culture as the gold standard, but these tests are generally expensive; depending on the manufacturer, require highly skilled molecular technologists; and have turnaround times of up to 24 h from receipt to results (1,7,(18)(19)(20)(21)(22). As a result, specimens with negative RIDTs are usually tested subsequently by more sensitive culture or molecular assays.…”

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confidence: 99%

Evaluation of Alere i Influenza A&B for Rapid Detection of Influenza Viruses A and B

Roth²,

et al. 2014

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Duplex Real-Time Reverse Transcriptase PCR Assays for Rapid Detection and Identification of Pandemic (H1N1) 2009 and Seasonal Influenza A/H1, A/H3, and B Viruses

Abstract: Reports of a novel influenza virus type A (H1N1),

Cited by 64 publications

References 10 publications

Viral Etiology and the Impact of Codetection in Young Children Presenting With Influenza-Like Illness

Viral Etiology and the Impact of Codetection in Young Children Presenting With Influenza-Like Illness

Development of a Multiplex Real-Time PCR Assay for the Detection of <i>Treponema pallidum</i>, HCV, HIV-1, and HBV

Evaluation of Alere i Influenza A&B for Rapid Detection of Influenza Viruses A and B

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