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1. Quantification of Legionella DNA certified reference material by digital droplet PCR

Author:
Baume, Maud; Cariou, Astrid; Leveau, Adélaïde; Fessy, Noémie; Pastori, Frédéric; Jarraud, Sophie; Pierre, Sophie
Source:
Journal of microbiological methods 2019 v.157 pp. 50-53
ISSN:
0167-7012
Subject:
quantitative polymerase chain reaction,   etc   ; DNA; Legionella; droplets;     Show all 4 Subjects
Abstract:
... A value was assigned in 2009 to the Legionella DNA Certified Reference Material, and the stability study conducted using quantitative PCR found a low level of degradation. Herein, the Digital Droplet PCR method for Legionella DNA was qualified and used to provide absolute quantification of the CRM. ...
DOI:
10.1016/j.mimet.2018.12.019

2. Droplet digital PCR for quantification of PML-RARα in acute promyelocytic leukemia: a comprehensive comparison with real-time PCR

Author:
Yuan, Dandan; Cui, Ming; Yu, Shuping; Wang, Huimin; Jing, Rongrong
Source:
Analytical and bioanalytical chemistry 2019 v.411 no.4 pp. 895-903
ISSN:
1618-2642
Subject:
quantitative polymerase chain reaction,   etc   ; detection limit; droplets; genes; leukemia; patients;     Show all 6 Subjects
Abstract:
... Real-time quantitative PCR (qPCR) has been widely implemented for molecular testing, but there are still some inherent limitations that hamper its usefulness. Droplet digital PCR (ddPCR), which can provide direct, standards-free quantification, has recently received increasing attention. In our study, a comprehensive comparison of ddPCR with qPCR in relation to the quantification of PML-RARα was p ...
DOI:
10.1007/s00216-018-1508-6

3. Comparative analysis of the performance of residual host cell DNA assays for viral vaccines produced in Vero cells

Author:
Vernay, Olivier; Sarcey, Eric; Detrez, Valérie; Abachin, Eric; Riou, Patrice; Mouterde, Bénédicte; Bonnevay, Thierry; Mallet, Laurent
Source:
Journal of virological methods 2019 v.268 pp. 9-16
ISSN:
0166-0934
Subject:
quantitative polymerase chain reaction,   etc   ; DNA; analytical methods; cell lines; hybridization; manufacturing; risk; viral vaccines;     Show all 8 Subjects
Abstract:
... Residual host cell DNA (rcDNA) from continuous cell lines used for manufacturing of biological medicinal products has been considered as safety risk. Historically, several analytical methods have been used for rcDNA quantitation including hybridization assay, Threshold® assay and quantitative polymerase chain reaction (qPCR). Sanofi Pasteur has a wealth of experience in the development of methods ...
DOI:
10.1016/j.jviromet.2019.01.001

4. Detection of phenol contamination in RNA samples and its impact on qRT-PCR results

Author:
Unger, Conny; Lokmer, Nicole; Lehmann, Daniel; Axmann, Ilka M.
Source:
Analytical biochemistry 2019 v.571 pp. 49-52
ISSN:
0003-2697
Subject:
quantitative polymerase chain reaction,   etc   ; RNA; absorbance; fluorometry; phenol; reverse transcriptase polymerase chain reaction;     Show all 6 Subjects
Abstract:
... Residual phenol, carried over from RNA purification, can alter RNA concentration measurements and is assumed to inhibit PCR. Here, we demonstrate that Impurities A260 values of spectral content profiling (SCP) UV/Vis measurements correlated with phenol concentration, whereas absorbance ratios of classical UV/Vis systems failed to reliably detect phenol in RNA samples. Phenol contamination led to o ...
DOI:
10.1016/j.ab.2019.02.002

5. MiR-486-5p inhibits the proliferation of leukemia cells and induces apoptosis through targeting FOXO1

Author:
Liu, Hui; Ni, Zengfeng; Shi, Lili; Ma, Lijie; Zhao, Jianqiang
Source:
Molecular and cellular probes 2019 v.44 pp. 37-43
ISSN:
0890-8508
Subject:
quantitative polymerase chain reaction,   etc   ; Western blotting; apoptosis; caspase-3; leukemia; leukocytes; neoplasm cells; therapeutics;     Show all 8 Subjects
Abstract:
... Studies have reported that micro (miR)-486-5p plays a crucial part in the progression of leukemia, however, to the best of our knowledge, few studies have been conducted on its mechanism in leukemia. In this study, the mechanism of miR-486-5p in leukemia cells was pointed out and its possible target genes were analyzed for the purpose of providing new therapeutic strategies for treating leukemia p ...
DOI:
10.1016/j.mcp.2019.02.001

6. Rapid start-up and stable maintenance of domestic wastewater nitritation through short-term hydroxylamine addition

Author:
Li, Jia; Zhang, Qiong; Li, Xiyao; Peng, Yongzhen
Source:
Bioresource technology 2019 v.278 pp. 468-472
ISSN:
0960-8524
Subject:
quantitative polymerase chain reaction,   etc   ; ammonia; bacteria; hydroxylamine; municipal wastewater; nitrification; nitrites;     Show all 7 Subjects
Abstract:
... This study investigated the nitritation of domestic wastewater through the short-term addition of hydroxylamine (NH2OH). Sequencing batch reactor (SBR) was used and the NH2OH solution with an initial concentration of 5.0 mg/L was injected at each cycle. With NH2OH addition, the nitritation was quickly established in 5 d with nitrite accumulation ratio above 95%. Further, stable nitritation could b ...
DOI:
10.1016/j.biortech.2019.01.056

7. Detection of goat meat adulteration by real-time PCR based on a reference primer

Author:
Li, Ting Ting; Jalbani, Yar Muhammad; Zhang, Gui Lan; Zhao, Zhi Yong; Wang, Zhi Ying; Zhao, Xiao Yan; Chen, Ai Liang
Source:
Food chemistry 2019 v.277 pp. 554-557
ISSN:
0308-8146
Subject:
quantitative polymerase chain reaction,   etc   ; adulterated products; food safety; goat meat; packaging; pork; quantitative analysis;     Show all 7 Subjects
Abstract:
... Economically motivated adulteration (EMA) has become one of the most concerned food safety issues. However, existing mainstream PCR methods could neither achieve qualitative detection purposes, nor detect all possible meat species involved in adulteration. When meat has been adulterated with unidentified species but the result indicates no adulteration, it is a false negative; when meat has not be ...
DOI:
10.1016/j.foodchem.2018.11.009

8. MicroRNA-150 promote apoptosis of ovine ovarian granulosa cells by targeting STAR gene

Author:
Zhou, Rongyan; Miao, Yanping; Li, Yimeng; Li, Xiangyun; Xi, Jianzhong; Zhang, Zhenhong
Source:
Theriogenology 2019 v.127 pp. 66-71
ISSN:
0093-691X
Subject:
quantitative polymerase chain reaction,   etc   ; apoptosis; estrous cycle; gene expression; genes; granulosa cells; luciferase; progesterone; sheep;     Show all 9 Subjects
Abstract:
... To better understand the function of oar-miR-150 in the ovine granulosa cells (GCs) during the estrus cycle, the five turpan sheep were selected for detection of the expression of oar-miR-150 in follicular and luteal ovaries, respectively. Then the granulosa cells treated with oar-miR-150 mimics or negative control (NC) were analyzed by qPCR to assess the expression of genes involved in steroidoge ...
DOI:
10.1016/j.theriogenology.2019.01.003

9. A novel real-time PCR assay panel for detection of common respiratory pathogens in a convenient, strip-tube array format

Author:
Hasan, Mohammad Rubayet; Al Mana, Hassan; Young, Virginia; Tang, Patrick; Thomas, Eva; Tan, Rusung; Tilley, Peter
Source:
Journal of virological methods 2019 v.265 pp. 42-48
ISSN:
0166-0934
Subject:
quantitative polymerase chain reaction,   etc   ; diagnostic techniques; microbial detection; nucleic acids; pathogens; respiratory tract diseases;     Show all 6 Subjects
Abstract:
... Commercial multiplex assays, built on different chemistries and platforms are widely available for simultaneous detection of pathogens that cause respiratory infections. However, these tests are often difficult to implement in a resource limited setting because of high cost. In this study, we developed and validated a method for simultaneous testing of common respiratory pathogens (Respanel) by re ...
DOI:
10.1016/j.jviromet.2018.12.013

10. Molecular detection, characterization of Anaplasma spp. in domestic cats from Rio de Janeiro state

Author:
Guimarães, Andresa; Raimundo, Juliana Macedo; Peixoto, Maristela Peckle; da Silva, Claudia Bezerra; Pires, Marcus Sandes; Santos, Huarrisson Azevedo; Baldani, Cristiane Divan
Source:
Acta tropica 2019 v.191 pp. 239-242
ISSN:
0001-706X
Subject:
quantitative polymerase chain reaction,   etc   ; Anaplasma phagocytophilum; Anaplasma platys; anaplasmosis; cats; microscopy; parasite load; Brazil;     Show all 8 Subjects
Abstract:
... Species of the genus Anaplasma, in the family Anaplasmatacae, are responsible to vector-borne diseases that affecting animals and humans. Feline anaplasmosis is poorly reported in Brazil. This study aimed at investigating the occurrence of Anaplasma spp. in domestic cats from Greater Rio de Janeiro, and evaluating hematological changes associated with this rickettsial infection. Were sampled 216 c ...
DOI:
10.1016/j.actatropica.2019.01.003

11. Evaluation of PCR-based methods for the identification of enteroaggregative hemorrhagic Escherichia coli in sprouts

Author:
Rotundo, Luca; Amagliani, Giulia; Carloni, Elisa; Omiccioli, Enrica; Magnani, Mauro; Paoli, George
Source:
International journal of food microbiology 2019 v.291 pp. 59-64
ISSN:
0168-1605
Subject:
quantitative polymerase chain reaction,   etc   ; Shiga toxin-producing Escherichia coli; agar; agglutination; alfalfa; genes; latex; mung beans;     Show all 8 Subjects
Abstract:
... In this study real-time PCR assays were evaluated for the detection of enteroaggregative hemorrhagic Escherichia coli (EAHEC) O104:H4 in artificially contaminated mung bean and/alfalfa sprouts inoculated with 1, 10, and 100 CFU of EAHEC O104:H4 per 25 g sample (20, 10, and 2 replicates respectively). After selective culture enrichment the samples were tested using commercial real-time PCR kits det ...
DOI:
10.1016/j.ijfoodmicro.2018.11.011

12. Two FAST multiplex real-time PCR reactions to assess the presence of genetically modified organisms in food

Author:
Cottenet, Geoffrey; Blancpain, Carine; Sonnard, Véronique; Chuah, Poh Fong
Source:
Food chemistry 2019 v.274 pp. 760-765
ISSN:
0308-8146
Subject:
quantitative polymerase chain reaction,   etc   ; biotechnology; business enterprises; detection limit; genetically modified organisms; laws and regulations; screening;     Show all 7 Subjects
Abstract:
... Methods to detect the presence of genetically modified organisms (GMOs) are evolving constantly to comply with legislation and include new transgenic traits developed by biotechnology companies. Since traditional screening methods target only a limited number of markers, not all GM-events can be detected easily. To cover a broader range, a new GMO screening method was developed, based on two real- ...
DOI:
10.1016/j.foodchem.2018.09.050

13. Towards dynamic monitoring of cell cultures using high throughput sequencing

Author:
McClenahan, Shasta D.; Krause, Philip R.
Source:
Vaccine 2019 v.37 no.7 pp. 1001-1005
ISSN:
0264-410X
Subject:
quantitative polymerase chain reaction,   etc   ; cell culture; cytopathogenicity; high-throughput nucleotide sequencing; monitoring; nucleic acids; vaccines; viruses;     Show all 8 Subjects
Abstract:
... We used a combination of DOP-PCR with high throughput sequencing (HTS) to study infected cell cultures over time to assess the feasibility of using this technique to provide a read-out other than cytopathic effect in cell culture infectivity assays. Because DOP-PCR primers feature a short constant sequence at their 3′ terminus, the procedure yields a reproducible representational library of produc ...
DOI:
10.1016/j.vaccine.2018.12.019

14. What counts? A review of in vitro methods for the enumeration of Mycobacterium avium subsp. paratuberculosis

Author:
Marquetoux, Nelly; Ridler, Anne; Heuer, Cord; Wilson, Peter
Source:
Veterinary microbiology 2019
ISSN:
0378-1135
Subject:
quantitative polymerase chain reaction,   etc   ; Mycobacterium avium subsp. paratuberculosis; in vitro studies; inoculum; paratuberculosis; turbidimetry; viability;     Show all 7 Subjects
Abstract:
... Enumeration of Mycobacterium avium subsp. paratuberculosis (MAP) from clinical samples or from inoculum suspensions for challenge studies is an essential pre-requisite for paratuberculosis research. However, MAP organisms are slow and challenging to grow in vitro, with a strong tendency to clump. Standard bacteriological methods are not well suited to enumerate MAP. Enumeration can be achieved by ...
DOI:
10.1016/j.vetmic.2019.02.011

15. Does vacuum centrifugal concentration reduce yield or quality of nucleic acids extracted from FFPE biospecimens?

Author:
Sánchez, Ignacio; Betsou, Fay; Mathieson, William
Source:
Analytical biochemistry 2019 v.566 pp. 16-19
ISSN:
0003-2697
Subject:
quantitative polymerase chain reaction,   etc   ; DNA; RNA; fluorescence emission spectroscopy; reverse transcriptase polymerase chain reaction; temperature;     Show all 6 Subjects
Abstract:
... Vacuum centrifugal (SpeedVac) concentration is commonly applied to nucleic acids extracted from formalin-fixed paraffin-embedded (FFPE) sections, but with an unknown impact. We concentrated DNA and RNA from FFPE biospecimens using different time/temperature SpeedVac combinations of up to 30 min concentration at 45 °C, then used spectrophotometry, spectrofluorometry, RIN, PERM, DV200, qRT-PCR, DIN ...
DOI:
10.1016/j.ab.2018.10.020

16. Development and application of a real-time polymerase chain reaction method for quantification of Escherichia coli in oysters (Crassostrea gigas)

Author:
Miotto, Marília; Fonseca, Antônio A.; Barretta, Clarissa; da Silva, Helen S.; Pellizzaro, Tomás; De Dea Lindner, Juliano; Vieira, Cleide R.W.; Parveen, Salina; Prudencio, Elane S.
Source:
Food microbiology 2019 v.77 pp. 85-92
ISSN:
0740-0020
Subject:
quantitative polymerase chain reaction,   etc   ; Crassostrea gigas; DNA; Escherichia coli; bacteria; genes; mariculture; oysters; pathogens; perishable foods;     Show all 10 Subjects
Abstract:
... Oysters are important mariculture species worldwide. Because of their filter-feeding behaviors, oysters can accumulate microorganisms, including pathogens, from surrounding water and concentrate bacteria in high numbers. Rapid and suitable methods for quantification of Escherichia coli in oysters are necessary considering that oysters are perishable foods often consumed raw and some countries use ...
DOI:
10.1016/j.fm.2018.08.015

17. High fidelity recovery of airborne microbial genetic materials by direct condensation capture into genomic preservatives

Author:
Nieto-Caballero, Marina; Savage, Nicole; Keady, Patricia; Hernandez, Mark
Source:
Journal of microbiological methods 2019 v.157 pp. 1-3
ISSN:
0167-7012
Subject:
quantitative polymerase chain reaction,   etc   ; air filters; bioaerosols; genome; genomics; microorganisms; microscopy; preservatives;     Show all 8 Subjects
Abstract:
... A condensation growth tube was adapted to capture bacterial bioaerosols directly into genomic preservatives. As judged by quantitative PCR and direct microscopy, bioaerosol condensation capture conserves airborne microbes' genomes as they exist in the atmospheric environment. This method circumvents the collection stresses bioaerosols experience on air filters, impactors and impingers. ...
DOI:
10.1016/j.mimet.2018.12.010

18. Human DNA degradation assessment and male DNA detection by quantitative-PCR followed by high-resolution melting analysis

Author:
Ginart, S.; Caputo, M.; Corach, D.; Sala, A.
Source:
Forensic science international 2019 v.295 pp. 1-7
ISSN:
0379-0738
Subject:
quantitative polymerase chain reaction,   etc   ; DNA; analytical kits; cost effectiveness; crime; females; humans; males; melting;     Show all 9 Subjects
Abstract:
... We developed a q-PCR technique that simultaneously evaluates the extent of degradation and determines the gender of a human DNA donor. QYDEG HRM is a triplex real-time PCR whose products are analysed by high-resolution melting (HRM). The system produces three amplicons: (1) transducin (beta)-like 1, Y-linked (TBL1Y) (84bp); (2) large-target sequence (DGlt) (244bp); and (3) small-target sequence (D ...
DOI:
10.1016/j.forsciint.2018.11.013

19. Improved detection of DNA Schistosoma haematobium from eggs extracted by bead beating in urine

Author:
Pomari, Elena; Perandin, Francesca; La Marca, Giulia; Bisoffi, Zeno
Source:
Parasitology research 2019 v.118 no.2 pp. 683-686
ISSN:
0932-0113
Subject:
quantitative polymerase chain reaction,   etc   ; DNA; Schistosoma haematobium; eggs; internal transcribed spacers; medicine; microscopy; protocols; urinalysis; urine;     Show all 10 Subjects
Abstract:
... Diagnosis of Schistosoma haematobium relies primarily on microscopical analysis of urine. The method is time consuming and requires some expertise. Genus-specific real-time PCRs have been developed, but we still observed low sensitivity. In the present study, in order to achieve a more sensitive DNA detection of eggs of S. haematobium in urine samples, we wanted to develop a novel protocol of DNA ...
DOI:
10.1007/s00436-018-6137-7

20. Patterns of expression of sperm and seminal plasma microRNAs in boar semen

Author:
Kasimanickam, Vanmathy; Buhr, Mary; Kasimanickam, Ramanathan
Source:
Theriogenology 2019 v.125 pp. 87-92
ISSN:
0093-691X
Subject:
quantitative polymerase chain reaction,   etc   ; boars; cell communication; complementary DNA; landraces; microRNA; seminal plasma; spermatozoa; testes;     Show all 9 Subjects
Abstract:
... Although sperm and seminal plasma differ in their origin, biophysical and biochemical properties of seminal plasma influence the sperm function. Seminal plasma is a fluid medium containing substances from testes, epididymides and accessory glands. Composition of seminal plasma varies among animal species and in boars, prostate and bulbourethral glands are major contributors to the volume and conte ...
DOI:
10.1016/j.theriogenology.2018.10.021