Detection of measles virus RNA on SYBR green real‐time reverse transcription–polymerase chain reaction

Ito, Masahiro; Suga, Tomoko; Akiyoshi, Kyoko; Nukuzuma, Souichi; Kon-no, Mayumi; Umegaki, Yasuhiro; Kohdera, Urara; Ihara, Toshiaki

doi:10.1111/j.1442-200x.2010.03124.x

Cited by 7 publications

(7 citation statements)

References 17 publications

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“…The thermocycling conditions were as follows: 37˚C for 15 min and 85˚C for 5 sec (for the reverse transcription); followed by 40 cycles of 95˚C for 5 sec and 60˚C for 34 sec. Relative fold changes were calculated using the 2 -∆∆Cq method (21) and standard curves were produced. The Cq values of the samples were normalized to the appropriate endogenous housekeeping gene, GAPDH.…”

Section: Effects Of Hydrogen Sulfide On the Expression Of Alkaline Phmentioning

confidence: 99%

Effects of hydrogen sulfide on the expression of alkaline phosphatase, osteocalcin and collagen type I in human periodontal ligament cells induced by tension force stimulation

Qin¹,

Hua²

2016

Molecular Medicine Reports

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Periodontal ligament cells (PDLCs) are important in homeostasis and remodeling in the mechanically‑stimulated periodontium. The aim of the present study was to investigate the effects of hydrogen sulfide (H2S) on periodontal tissue remodeling by examining the mRNA and protein expression levels of alkaline phosphatase (ALP), osteocalcin (OCN) and collagen type I (COL‑1) in human (h)PDLCs induced by tension force application. Cultured hPDLCs were treated with H2S for 24 h, followed by application of a tension force for 1, 3 and 6 h. Cell proliferation and apoptosis were determined using a Cell Counting Kit 8 assay and flow cytometric analysis, respectively. The mRNA expression levels of ALP, OCN and COL‑1 were quantified using reverse transcription‑quantitative polymerase chain reaction analysis, and western blot analysis was used to detect the protein levels of ALP, OCN and COL‑1. The results demonstrated that the mRNA and protein expression levels of ALP, OCN and COL‑1 increased with H2S treatment in a concentration‑dependent manner, which was enhanced by the application of tension force in a relatively short period of time. These findings suggested that H2S may be important in periodontal tissue remodeling during orthodontic tooth movement via increasing hPDLC differentiation, tissue mineralization, bone formation and collagen synthesis.

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Section: Effects Of Hydrogen Sulfide On the Expression Of Alkaline Phmentioning

confidence: 99%

Effects of hydrogen sulfide on the expression of alkaline phosphatase, osteocalcin and collagen type I in human periodontal ligament cells induced by tension force stimulation

Qin¹,

Hua²

2016

Molecular Medicine Reports

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“…Reverse transcription polymerase chain reaction is a sensitive tool for detecting influenza virus, but this method is unable to quantitatively detect viral loads. We have already reported that real‐time RT‐PCR is a sensitive, rapid, and useful diagnostic procedure for the detection of measles virus RNA 6 . In the present study, we evaluated copy number of pandemic H1N1 virus RNA in nasopharyngeal aspirates using real‐time RT‐PCR.…”

Section: Discussionmentioning

confidence: 92%

“…We have already reported that real-time RT-PCR is a sensitive, rapid, and useful diagnostic procedure for the detection of measles virus RNA. 6 In the present study, we evaluated copy number of pandemic H1N1 virus RNA in nasopharyngeal aspirates using real-time RT-PCR. Studies on changes in viral load in pandemic H1N1 influenza patients using real-time RT-PCR have been reported previously.…”

Section: Discussionmentioning

confidence: 99%

Viral load and rapid diagnostic test in patients with pandemic H1N1 2009

Watanabe

Nukuzuma²,

Ito

et al. 2011

Pediatrics International

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“…Methods used for laboratory confirmation are affected by the timing of specimen collection [ 24 – 26 ]. In fact, studies have shown that 30% of serum specimens obtained in the first 72 hours after rash onset reveal negative results, since IgM antibodies are still developing and may be below detectable levels[ 22 , 27 , 28 ]. Taking into account that case investigation and sampling, in our context, occur early after rash onset, this implies that the number of measles cases reported maybe slightly underestimated based on IgM serology.…”

Section: Discussionmentioning

confidence: 99%

Real-Time PCR for Measles Virus Detection on Clinical Specimens with Negative IgM Result in Morocco

et al. 2016

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Since the confirmation of measles cases represents an important indicator regarding the performance of the measles-elimination program, the aim of this study was to evaluate the effectiveness of the routine procedures followed in Morocco for the laboratory confirmation of measles cases. Suspected cases reported between January 2010 and December 2012 were assessed for the timeliness of the sample collection, occurrence of measles clinical symptoms, and the results of the laboratory diagnoses. For 88% of the 2,708 suspected cases, a clinical specimen was collected within 7d of rash onset, of which 50% were IgM-positive and 2.6% were equivocal. The measles symptoms were reported in 91.4% of the cases; the occurrence of symptoms showed a positive association with the serological results (odds ratio [OR] = 2.9883, 95% confidence interval [CI] 2.2238–4.0157). Of the negative samples, 52% (n = 116) tested positive by real-time polymerase chain reaction (PCR). These results are in favor of using molecular detection to complement serological diagnosis in the context of measles surveillance approach in Morocco. In addition, the introduction of additional laboratory methods for differential diagnosis is required for the final classification of suspected cases with maculopapular rash and fever in the context of the measles elimination program.

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Detection of measles virus RNA on SYBR green real‐time reverse transcription–polymerase chain reaction

Abstract: SYBR Green RT-PCR is a highly sensitive, rapid, and useful diagnostic procedure for the detection of MV.

Cited by 7 publications

References 17 publications

Effects of hydrogen sulfide on the expression of alkaline phosphatase, osteocalcin and collagen type I in human periodontal ligament cells induced by tension force stimulation

Effects of hydrogen sulfide on the expression of alkaline phosphatase, osteocalcin and collagen type I in human periodontal ligament cells induced by tension force stimulation

Viral load and rapid diagnostic test in patients with pandemic H1N1 2009

Real-Time PCR for Measles Virus Detection on Clinical Specimens with Negative IgM Result in Morocco

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