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Rapid PCR

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https://www.readbyqxmd.com/read/28101989/diagnosis-of-peste-des-petits-ruminants-in-wild-and-domestic-animals-in-xinjiang-china-2013-2016
#1
J Li, L Li, X Wu, F Liu, Y Zou, Q Wang, C Liu, J Bao, W Wang, W Ma, H Lin, J Huang, X Zheng, Z Wang
Peste des petits ruminants viruses (PPRVs) re-emerged in China at the end of 2013 and then spread rapidly into 22 provinces through movement of live goats and sheep. In this study, 96 samples of domestic animals and 13 samples of wildlife were analysed for the presence of PPRV infection by ELISA or RT-PCR. Of 96 samples from sheep and goats, 91 were PPRV positive, whereas all of the 13 samples from three wild species, Capra ibex (Capra ibex sibirica), argali (Ovis ammon) and Goitered gazelle (Gazella subgutturosa), were found to be positive...
January 18, 2017: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/28100599/rapid-and-specific-detection-of-the-escherichia-coli-sequence-type-648-complex-stc648-within-phylogroup-f
#2
James R Johnson, Brian D Johnston, David M Gordon
The Escherichia coli sequence type 648 complex (STc648) is an emerging lineage within phylogroup F - formerly included within phylogroup D - that is associated with multidrug resistance. Here, we designed and validated a novel multiplex PCR-based assay for STc648 that took advantage of (i) four distinctive single-nucleotide polymorphisms in icd allele 96 and gyrB allele 87, two of the multi-locus sequence typing alleles that define ST648, and (ii) the typical absence within STc648 of uidA, an E. coli-specific gene encoding β-glucuronidase...
January 18, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28100259/antitumor-activity-of-mir-34a-in-peritoneal-mesothelioma-relies-on-c-met-and-axl-inhibition-persistent-activation-of-erk-and-akt-signaling-as-a-possible-cytoprotective-mechanism
#3
Rihan El Bezawy, Michelandrea De Cesare, Marzia Pennati, Marcello Deraco, Paolo Gandellini, Valentina Zuco, Nadia Zaffaroni
BACKGROUND: The value of microRNAs (miRNAs) as novel targets for cancer therapy is now widely recognized. However, no information is currently available on the expression/functional role of miRNAs in diffuse malignant peritoneal mesothelioma (DMPM), a rapidly lethal disease, poorly responsive to conventional treatments, for which the development of new therapeutic strategies is urgently needed. Here, we evaluated the expression and biological effects of miR-34a-one of the most widely deregulated miRNAs in cancer and for which a lipid-formulated mimic is already clinically available-in a large cohort of DMPM clinical samples and a unique collection of in house-developed preclinical models, with the aim to assess the potential of a miR-34a-based approach for disease treatment...
January 18, 2017: Journal of Hematology & Oncology
https://www.readbyqxmd.com/read/28099753/correlation-between-rapid-hiv-testing-and-fourth-generation-elisa-results-for-hiv-detection-among-pregnant-patients-in-the-delivery-room
#4
Alejandro G Almaguer, Lidia Mendoza-Flores, Luis A Sánchez-López, Laura A Palau-Dávila, Magaly Padilla-Orozco, Adrián Camacho-Ortiz
OBJECTIVE: To analyze the usefulness of rapid HIV testing in pregnant patients in the delivery room. METHODS: This prospective study compared a rapid test and a fourth-generation enzyme-linked immunoassay (ELISA) for HIV screening among pregnant patients admitted in labor with an unknown HIV status at a university hospital in Mexico between July 2015 and February 2016. Pearson correlation analysis was performed, and the diagnostic accuracy of the two tests was assessed with HIV RNA polymerase chain reaction (PCR) as the reference method...
January 5, 2017: International Journal of Gynaecology and Obstetrics
https://www.readbyqxmd.com/read/28099512/age-dependent-oxidative-dna-damage-does-not-correlate-with-reduced-proliferation-of-cardiomyocytes-in-humans
#5
Yanhui Huang, Haifa Hong, Minghui Li, Jinfen Liu, Chuan Jiang, Haibo Zhang, Lincai Ye, Jinghao Zheng
BACKGROUND: Postnatal human cardiomyocyte proliferation declines rapidly with age, which has been suggested to be correlated with increases in oxidative DNA damage in mice and plays an important role in regulating cardiomyocyte proliferation. However, the relationship between oxidative DNA damage and age in humans is unclear. METHODS: Sixty right ventricular outflow myocardial tissue specimens were obtained from ventricular septal defect infant patients during routine congenital cardiac surgery...
2017: PloS One
https://www.readbyqxmd.com/read/28098507/development-and-evaluation-of-a-loop-mediated-isothermal-amplification-lamp-assay-for-the-detection-of-haemonchus-contortus-in-goat-fecal-samples
#6
Xin Yang, Mingwei Qi, Zongze Zhang, Chong Gao, Chunqun Wang, Weiqiang Lei, Li Tan, Junlong Zhao, Rui Fang, Min Hu
Haemonchus contortus is one of the most significant strongylid nematodes infecting small ruminants and causes great economic losses to the livestock industry worldwide. Accurate diagnosis of H. contortus is crucial to control the infection. Traditional microscopic examinations are the most common methods for the diagnosis of H. contortus, but they are time-consuming and inaccurate. Molecular methods based on PCR are more accurate, but need expensive machines usually only used in the laboratory. Loop-mediated isothermal amplification (LAMP) is a rapid, simple, specific and sensitive method that has been widely used to detect viruses, bacteria and parasites...
January 18, 2017: Journal of Parasitology
https://www.readbyqxmd.com/read/28097865/molecular-requirements-of-high-fidelity-replication-competent-dna-backbones-for-orthogonal-chemical-ligation
#7
Arun Shivalingam, Agnes E S Tyburn, Afaf H El-Sagheer, Tom Brown
The molecular properties of the phosphodiester backbone that made it the evolutionary choice for the enzymatic replication of genetic information are not well understood. To address this, and to develop new chemical ligation strategies for assembly of biocompatible modified DNA, we have synthesized oligonucleotides containing several structurally and electronically varied artificial linkages. This has yielded a new highly promising ligation method based on amide backbone formation that is chemically orthogonal to CuAAC "click" ligation...
January 18, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28096860/determination-of-asymptomatic-malaria-among-afghani-and-pakistani-immigrants-and-native-population-in-south-of-kerman-province-iran
#8
Mohammad Bagher Amirshekari, Mehdi Nateghpour, Ahmad Raeisi, Afsaneh Motevalli Haghi, Leila Farivar, Gholamhosein Edrissian
BACKGROUND: This study was proposed to monitor the situation of asymptomatic malaria among the native population and Afghani and Pakistani immigrants in Kahnooj and Ghale-Ganj districts from Kerman Province, Southeastern Iran. METHODS: A number of 180 and 120 individuals from Kahnooj and Ghale-Ganj respectively were registered and considered based on a cross-sectional surveillance method. From 300 registered cases, 200 individuals (66.7%) were selected among Afghani and Pakistani immigrants and the rest (33...
April 2016: Iranian Journal of Parasitology
https://www.readbyqxmd.com/read/28095801/providing-lipid-based-nutrient-supplement-during-pregnancy-does-not-reduce-the-risk-of-maternal-p-falciparum-parasitaemia-and-reproductive-tract-infections-a-randomised-controlled-trial
#9
Minyanga Nkhoma, Per Ashorn, Ulla Ashorn, Kathryn G Dewey, Austrida Gondwe, John Mbotwa, Stephen Rogerson, Steve M Taylor, Kenneth Maleta
BACKGROUND: Maternal infections are associated with maternal and foetal adverse outcomes. Nutrient supplementation during pregnancy may reduce the occurrence of infections by improving maternal immunity. We aimed to investigate the impact of small-quantity lipid-based nutrient supplement (SQ-LNS) on the occurrence of Plasmodium falciparum parasitaemia during pregnancy and trichomoniasis, vaginal candidiasis and urinary tract infection (UTI) after delivery. METHODS: Pregnant Malawian women enrolled in the iLiNS-DYAD trial receiving daily supplementation with SQ-LNS, multiple micronutrients (MMN) or iron & folic acid (IFA) from <20 gestation weeks (gw) were assessed for P...
January 17, 2017: BMC Pregnancy and Childbirth
https://www.readbyqxmd.com/read/28095683/colorimetric-detection-of-staphylococcus-aureus-contaminated-solutions-without-purification
#10
Pamela Tiet, Karen C Clark, James O McNamara, Jacob M Berlin
Current water quality monitoring methods rely on growth-based measurements to detect fecal indicator bacteria, such as Escherichia coli and enterococci, and Staphylococcus aureus (S. aureus). These growth-based measurements, however, can take days to complete. This is a significant limitation in the evaluation of contaminated food and water sources. Various methods for selective in vitro detection of S. aureus have also been reported; however, these strategies, such as ELISA, agar-diffusion, PCR, or liquid chromatography-tandem mass spectrometry, all require overnight culturing or sophisticated instrumentation...
January 18, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28095454/non-homologous-end-joining-and-homology-directed-dna-repair-frequency-of-double-stranded-breaks-introduced-by-genome-editing-reagents
#11
Michail Zaboikin, Tatiana Zaboikina, Carl Freter, Narasimhachar Srinivasakumar
Genome editing using transcription-activator like effector nucleases or RNA guided nucleases allows one to precisely engineer desired changes within a given target sequence. The genome editing reagents introduce double stranded breaks (DSBs) at the target site which can then undergo DNA repair by non-homologous end joining (NHEJ) or homology directed recombination (HDR) when a template DNA molecule is available. NHEJ repair results in indel mutations at the target site. As PCR amplified products from mutant target regions are likely to exhibit different melting profiles than PCR products amplified from wild type target region, we designed a high resolution melting analysis (HRMA) for rapid identification of efficient genome editing reagents...
2017: PloS One
https://www.readbyqxmd.com/read/28095447/identification-and-characterization-of-single-chain-antibodies-that-specifically-bind-gi-noroviruses
#12
Amy M Hurwitz, Wanzhi Huang, Baijun Kou, Mary K Estes, Robert L Atmar, Timothy Palzkill
Norovirus infections commonly lead to outbreaks of acute gastroenteritis and spread quickly, resulting in many health and economic challenges prior to diagnosis. Rapid and reliable diagnostic tests are therefore essential to identify infections and to guide the appropriate clinical responses at the point-of-care. Existing tools, including RT-PCR and enzyme immunoassays, pose several limitations based on the significant time, equipment and expertise required to elicit results. Immunochromatographic assays available for use at the point-of-care have poor sensitivity and specificity, especially for genogroup I noroviruses, thus requiring confirmation of results with more sensitive testing methods...
2017: PloS One
https://www.readbyqxmd.com/read/28094255/specific-and-sensitive-isothermal-electrochemical-biosensor-for-plant-pathogen-dna-detection-with-colloidal-gold-nanoparticles-as-probes
#13
Han Yih Lau, Haoqi Wu, Eugene J H Wee, Matt Trau, Yuling Wang, Jose R Botella
Developing quick and sensitive molecular diagnostics for plant pathogen detection is challenging. Herein, a nanoparticle based electrochemical biosensor was developed for rapid and sensitive detection of plant pathogen DNA on disposable screen-printed carbon electrodes. This 60 min assay relied on the rapid isothermal amplification of target pathogen DNA sequences by recombinase polymerase amplification (RPA) followed by gold nanoparticle-based electrochemical assessment with differential pulse voltammetry (DPV)...
January 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28093921/microbial-genomics-and-antimicrobial-susceptibility-testing
#14
W Michael Dunne, Magali Jaillard, Olivier Rochas, Alex van Belkum
Antimicrobial susceptibility testing is key in modern clinical microbiology. With pandemic emergence of (multi-)antibiotic resistance, methods to detect and quantify resistance of clinically important bacterial species are imperative. Historically, antimicrobial susceptibility testing (AST) was mostly performed using methods relying on bacterial growth. Such methods may be time-consuming and more rapid alternatives have been actively sought for. Areas covered: Among the new AST methods there are many that focus on detection of causal resistance genes and/or gene mutations...
January 17, 2017: Expert Review of Molecular Diagnostics
https://www.readbyqxmd.com/read/28093275/reverse-transcription-loop-mediated-isothermal-amplification-for-rapid-detection-of-respiratory-syncytial-virus-directly-from-nasopharyngeal-swabs
#15
Johannes Hoos, Rebecca Marie Peters, Julia Tabatabai, Jürgen Grulich-Henn, Paul Schnitzler, Johannes Pfeil
Respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infection in young infants and a major cause of nosocomial infection in pediatric care. Currently available RSV point-of-care tests are of limited sensitivity and relatively expensive. We developed and evaluated a novel RSV rapid test for use at point-of-care, based on reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for direct testing of nasopharyngeal swab specimens. RT-LAMP can detect RSV within 30min, without the need for RNA extraction...
January 13, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28092867/expression-of-mmp-2-and-timp-1-during-rapid-maxillary-expansion-in-rats
#16
Jianwei Chen, Jing Zhou, Fan Li, Jianfeng Sun, Guifeng Li, Shujuan Zou, Qingsong Ye
OBJECTIVE: The aim of this study was to investigate the expression of MMP-2 and TIMP-1 during midpalatal suture expansion in rats. DESIGN: 72 male Wistar rats were randomly divided into 2 groups: the experimental group and the control group. In the experimental group, opening loops were applied across the midpalatal suture with an initial force of 50g, whereas in the control group, rats were subjected to sham installation of opening loops without activation. On day 1, 4, 7 and 14, nine rats from each group were sacrificed, and the maxillae were dissected and prepared for Immunohistochemistry (IHC) and RT- PCR examination of MMP-2 and TIMP-1 expression...
January 4, 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/28092041/in-solution-sh2-domain-binding-assay-based-on-proximity-ligation
#17
Kazuya Machida
Protein-protein interactions mediated by SH2 domains confer specificity in tyrosine kinase pathways. Traditional assays for assessing interactions between an SH2 domain and its interacting protein such as far-Western and pull-down are inherently low throughput. We developed SH2-PLA, an in-solution SH2 domain binding assay, that takes advantage of the speed and sensitivity of proximity ligation and real-time PCR. SH2-PLA allows for rapid assessment of SH2 domain binding to a target protein using only a few microliters of cell lysate, thereby making it an attractive new tool to study tyrosine kinase signaling...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28089896/characterization-and-expression-analysis-of-a-chitinase-gene-pmchi-4-from-black-tiger-shrimp-penaeus-monodon-under-pathogen-infection-and-ambient-ammonia-nitrogen-stress
#18
Kaimin Zhou, Falin Zhou, Jianhua Huang, Qibin Yang, Song Jiang, Lihua Qiu, Lishi Yang, Caiyan Zhu, Shigui Jiang
Chitinase is a multi-gene family, which play important physiological roles in crustaceans, involved in several biological processes, including digestion, molting and defense against viruses. In the present study, a chitinase-4 gene (PmChi-4) was cloned from Penaeus monodon by rapid amplification of cDNA ends (RACE). The full length of PmChi-4 cDNA was 2178 bp, including an 1815 bp open reading frame (ORF) which encoded 604 amino acid residues. The predicted PmChi-4 protein was 67.7 kDa and shared 61%-88% identity with the type of Chi-4s from other crustaceans...
January 9, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28089246/identification-and-characterization-of-l-lysine-decarboxylase-from-huperzia-serrata-and-its-role-in-the-metabolic-pathway-of-lycopodium-alkaloid
#19
Baofu Xu, Lei Lei, Xiaocen Zhu, Yiqing Zhou, Youli Xiao
Lysine decarboxylation is the first biosynthetic step of Huperzine A (HupA). Six cDNAs encoding lysine decarboxylases (LDCs) were cloned from Huperzia serrata by degenerate PCR and rapid amplification of cDNA ends (RACE). One HsLDC isoform was functionally characterized as lysine decarboxylase. The HsLDC exhibited greatest catalytic efficiency (kcat/Km, 2.11 s(-1) mM(-1)) toward L-lysine in vitro among all reported plant-LDCs. Moreover, transient expression of the HsLDC in tobacco leaves specifically increased cadaverine content from zero to 0...
January 12, 2017: Phytochemistry
https://www.readbyqxmd.com/read/28088897/multiplex-pcr-as-a-rapid-and-sensitive-method-for-identification-of-meat-species-in-halal-meat-products
#20
Mahsa Alikord, Javad Keramat, Mahdi Kadivar, Hassan Momtaz, Mohammad Naghi Eshtiaghi, Aziz Homayouni Rad
BACKGROUND: Species identification and authentication in meat products are important subjects for ensuring the health of consumers. The multiplex-PCR amplification and species-specific primer set were used for identification of horse, donkey, pig and other ruminants in raw and processed meat products. METHODS: Oligonucleotid primers were designed for amplification of species-specific mitochondrial DNA sequences of each species and samples were prepared from binary meat mixtures...
January 13, 2017: Recent Patents on Food, Nutrition & Agriculture
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