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Multiplex PCR

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https://www.readbyqxmd.com/read/28646669/validation-of-a-commercial-kit-aimed-to-the-detection-of-pathogenic-anisakid-nematodes-in-fish-products
#1
Serena Cavallero, Alessandro Bruno, Enrico Arletti, Monica Caffara, Maria Letizia Fioravanti, Antonella Costa, Gaetano Cammilleri, Stefania Graci, Vincenzo Ferrantelli, Stefano D'Amelio
Anisakids are parasitic nematodes responsible for a zoonosis that occurs following the ingestion of fish and fish products infected with larvae belonging to the genera Anisakis and Pseudoterranova. Rarely Contracaecum is found in association with gastric/intestinal illness, while Hysterothylacium is commonly considered not pathogenic. Although Real Time PCR assays have been recently used with the aim to detect and quantify these parasites in food products, methods applied did not undergo through extensive validation process, a feature highly desirable or mandatory in the case of testing laboratories accredited for the ISO EN 17025:2005...
June 15, 2017: International Journal of Food Microbiology
https://www.readbyqxmd.com/read/28643492/human-papillomavirus-genotype-distribution-among-18-815-women-in-13-korean-cities-and-relationship-with-cervical-cytology-findings
#2
Eun Hee Nah, Seon Cho, Suyoung Kim, Han Ik Cho
BACKGROUND: The prevalence and genotype distribution of Human papillomavirus (HPV) infection vary depending on geographical region and the immunity provided by vaccines. This study aimed to clarify the recent prevalence and genotype distribution of HPV according to age and cervical cytology findings in Korea. METHODS: This study included 18,815 health examinees that underwent cervical cytology and HPV genotyping tests at 16 centers of Korean Association of Health Promotion in 13 cities in Korea, between January 2014 and October 2015...
September 2017: Annals of Laboratory Medicine
https://www.readbyqxmd.com/read/28643489/comparison-of-the-luminex-xtag-respiratory-viral-panel-fast-v2-assay-with-anyplex-ii-rv16-detection-kit-and-advansure-rv-real-time-rt-pcr-assay-for-the-detection-of-respiratory-viruses
#3
Dae Hyun Ko, Hyun Soo Kim, Jungwon Hyun, Han Sung Kim, Jae Seok Kim, Kyoung Un Park, Wonkeun Song
BACKGROUND: The accurate and rapid identification of the causative viruses is important for the timely diagnosis and management of respiratory infections. Multiplex molecular diagnostic techniques have been widely adopted to detect respiratory viruses. We compared the results of a newly upgraded, multiplex, molecular bead-based respiratory viral panel (RVP) assay with the results of Anyplex II RV16 detection kit and AdvanSure RV real-time RT-PCR assay. METHODS: We tested 254 respiratory specimens and cultured viral strains using the Luminex xTAG RVP Fast v2 assay (Luminex Molecular Diagnostics, Canada) and Anyplex II RV16 detection kit and compared the results...
September 2017: Annals of Laboratory Medicine
https://www.readbyqxmd.com/read/28642764/xylella-fastidiosa-host-range-and-advance-in-molecular-identification-techniques
#4
REVIEW
Paolo Baldi, Nicola La Porta
In the never ending struggle against plant pathogenic bacteria, a major goal is the early identification and classification of infecting microorganisms. Xylella fastidiosa, a Gram-negative bacterium belonging to the family Xanthmonadaceae, is no exception as this pathogen showed a broad range of vectors and host plants, many of which may carry the pathogen for a long time without showing any symptom. Till the last years, most of the diseases caused by X. fastidiosa have been reported from North and South America, but recently a widespread infection of olive quick decline syndrome caused by this fastidious pathogen appeared in Apulia (south-eastern Italy), and several cases of X...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28639594/viral-aetiology-of-wheezing-in-children-under-five
#5
Prithi Sureka Mummidi, Radha Tripathy, Bhagirathi Dwibedi, Amarendra Mahapatra, Suryakanta Baraha
BACKGROUND & OBJECTIVES: Wheezing is a common problem in children under five with acute respiratory infections (ARIs). Viruses are known to be responsible for a considerable proportion of ARIs in children. This study was undertaken to know the viral aetiology of wheezing among the children less than five years of age, admitted to a tertiary care hospital in eastern India. METHODS: Seventy five children, under the age of five years admitted with wheezing, were included in the study...
February 2017: Indian Journal of Medical Research
https://www.readbyqxmd.com/read/28638437/a-multiplex-pcr-for-rapid-identification-of-brassica-species-in-the-triangle-of-u
#6
Joshua C O Koh, Denise M Barbulescu, Sally Norton, Bob Redden, Phil A Salisbury, Sukhjiwan Kaur, Noel Cogan, Anthony T Slater
BACKGROUND: Within the Brassicaceae, six species from the genus Brassica are widely cultivated throughout the world as oilseed, condiment, fodder or vegetable crops. The genetic relationships among the six Brassica species are described by U's triangle model. Extensive shared traits and diverse morphotypes among Brassica species make identification and classification based on phenotypic data alone challenging and unreliable, especially when dealing with large germplasm collections. Consequently, a major issue for genebank collections is ensuring the correct identification of species...
2017: Plant Methods
https://www.readbyqxmd.com/read/28638416/corrigendum-to-detection-of-gastrointestinal-pathogens-from-stool-samples-on-hemoccult-cards-by-multiplex-pcr
#7
Martin Alberer, Nicklas Schlenker, Malkin Bauer, Kerstin Helfrich, Carolin Mengele, Thomas Löscher, Hans Dieter Nothdurft, Gisela Bretzel, Marcus Beissner
[This corrects the article DOI: 10.1155/2017/3472537.].
2017: Canadian Journal of Infectious Diseases & Medical Microbiology
https://www.readbyqxmd.com/read/28637945/multiplex-pcr-assay-for-the-simultaneous-detection-and-differentiation-of-clonal-lineages-of-erysipelothrix-rhusiopathiae-serovar-1a-strains-currently-circulating-in-japan
#8
Kazumasa Shiraiwa, Yohsuke Ogawa, Sayaka Nishikawa, Masahiro Eguchi, Yoshihiro Shimoji
The species Erysipelothrix rhusiopathiae displays genetic heterogeneity; however, E. rhusiopathiae serovar 1a strains currently circulating in Japan exhibit remarkably low levels of genetic diversity and group into clonal sublineages of Lineage IVb (IVb-1 and IVb-2). In the present study, based on whole genome sequencing data, we designed primers for a multiplex PCR assay to simultaneously detect and differentiate the sublineages of E. rhusiopathiae strains. Among the one hundred and twenty-seven isolates of various serovar strains, including isolates from a wide range of hosts and geographic origins, the PCR assay could successfully detect and differentiate the serovar 1a strains belonging to the sublineages...
June 22, 2017: Journal of Veterinary Medical Science
https://www.readbyqxmd.com/read/28636980/a-multiplex-liquid-chip-assay-based-on-luminex-xmap-technology-for-simultaneous-detection-of-six-common-respiratory-viruses
#9
Yong Yan, Jian-Yong Luo, Yin Chen, Heng-Hui Wang, Guo-Ying Zhu, Pei-Yan He, Jin-Lei Guo, Yong-Liang Lei, Zhong-Wen Chen
We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR...
June 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/28636636/sensiscreen%C3%A2-kras-exon-2-sensitive-simplex-and-multiplex-real-time-pcr-based-assays-for-detection-of-kras-exon-2-mutations
#10
Alice Riva, Michael BØrgesen, Mariann Guldmann-Christensen, Majbritt Hauge Kyneb, Kirsten Voogd, Christina Andersen, Samantha Epistolio, Elisabetta Merlo, Tine Yding Wolff, Stephen Hamilton-Dutoit, Jan Lorenzen, Ulf Bech Christensen, Milo Frattini
Activating mutations in codon 12 and codon 13 of the KRAS (Kirsten rat sarcoma viral oncogene homolog) gene are implicated in the development of several human cancer types and influence their clinical evaluation, treatment and prognosis. Numerous different methods for KRAS genotyping are currently available displaying a wide range of sensitivities, time to answer and requirements for laboratory equipment and user skills. Here we present SensiScreen® KRAS exon 2 simplex and multiplex CE IVD assays, that use a novel real-time PCR-based method for KRAS mutation detection based on PentaBase's proprietary DNA analogue technology and designed to work on standard real-time PCR instruments...
2017: PloS One
https://www.readbyqxmd.com/read/28636544/analysis-of-esr1-and-pik3ca-mutations-in-plasma-cell-free-dna-from-er-positive-breast-cancer-patients
#11
Takashi Takeshita, Yutaka Yamamoto, Mutsuko Yamamoto-Ibusuki, Mai Tomiguchi, Aiko Sueta, Keiichi Murakami, Yoko Omoto, Hirotaka Iwase
BACKGROUND: The measurement of ESR1 and PIK3CA mutations in plasma cell-free DNA (cfDNA) has been studied as a non-invasive method to quickly assess and monitor endocrine therapy (ET) resistant metastatic breast cancer (MBC) patients. METHODS: The subjects of this retrospective study were a total of 185 plasma samples from 86 estrogen receptor-positive BC patients, of which 151 plasma samples were from 69 MBC patients and 34 plasma samples were from 17 primary BC (PBC) patients...
June 14, 2017: Oncotarget
https://www.readbyqxmd.com/read/28634689/survival-intestinal-mucosa-adhesion-and-immunomodulatory-potential-of-lactobacillus-plantarum-strains
#12
Valentini Santarmaki, Yiannis Kourkoutas, Georgia Zoumpopoulou, Eleni Mavrogonatou, Mikis Kiourtzidis, Nikos Chorianopoulos, Chrysoula Tassou, Effie Tsakalidou, Constantinos Simopoulos, Petros Ypsilantis
Survival during transit through the gastrointestinal track, intestinal mucosa adhesion, and a potential immunomodulatory effect of Lactobacillus plantarum strains 2035 and ACA-DC 2640 were investigated in a rat model. According to microbiological and multiplex PCR analysis, both strains were detected in feces 24 h after either single-dose or daily administration for 7 days. Intestinal mucosa adhesion of L. plantarum 2035 was noted in the large intestine at 24 h after single-dose administration, while it was not detected at 48 h...
June 21, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28634597/molecular-characterization-of-salmonella-from-human-and-animal-origins-in-uganda
#13
Atek Atwiine Kagirita, Andrew Baguma, Tonny Jimmy Owalla, Joel Bazira, Samuel Majalija
Sporadic Salmonella outbreaks with varying clinical presentations have been on the rise in various parts of Uganda. The sources of outbreaks and factors underlying the different clinical manifestation are curtailed by paucity of information on Salmonella genotypes and the associated virulence genes. This study reports molecular diversity of Salmonella enterica and their genetic virulence profiles among human and animal isolates. Characterization was done using Kauffman-White classification scheme and virulence genes analysis using multiplex PCR...
2017: International Journal of Bacteriology
https://www.readbyqxmd.com/read/28634144/development-of-a-multiplex-pcr-assay-for-the-detection-and-differentiation-of-burkholderia-pseudomallei-burkholderia-mallei-burkholderia-thailandensis-and-burkholderia-cepacia-complex
#14
Irina Zakharova, Natalya Teteryatnikova, Andrey Toporkov, Dmitry Viktorov
Two species of Burkholderia pseudomallei complex (Bpc), B. pseudomallei and B. mallei, can cause severe life-threatening infections. Rapidly discerning individual species within the group and separating them from other opportunistic pathogens of the Burkholderia cepacia complex (Bcc) is essential to establish a correct diagnosis and for epidemiological surveillance. In this study, a multiplex PCR assay based on the detection of an individual set of chromosomal beta-lactamase genes for single-step identification and differentiation of B...
June 17, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28634143/pcr-based-verification-of-positive-rapid-diagnostic-tests-for-intestinal-protozoa-infections-with-variable-test-band-intensity
#15
Sören L Becker, Ivan Müller, Pascal Mertens, Mathias Herrmann, Leyli Zondie, Lindsey Beyleveld, Markus Gerber, Rosa du Randt, Uwe Pühse, Cheryl Walter, Jürg Utzinger
Stool-based rapid diagnostic tests (RDTs) for pathogenic intestinal protozoa (e.g. Cryptosporidium spp. and Giardia intestinalis) allow for prompt diagnosis and treatment in resource-constrained settings. Such RDTs can improve individual patient management and facilitate population-based screening programmes in areas without microbiological laboratories for confirmatory testing. However, RDTs are difficult to interpret in case of 'trace' results with faint test band intensities and little is known about whether such ambiguous results might indicate 'true' infections...
June 17, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28633962/evaluation-of-three-commercial-multiplex-assays-for-the-detection-of-respiratory-viral-infections
#16
Sandi Radko, J Ian Stuart, George Zahariadis
BACKGROUND: Timely identification of respiratory virus infection is essential to mitigate inappropriate antibiotic use and to implement appropriate treatment and/or infection control procedures. As such, multiplexed PCR assays have become standard in many virology laboratories. OBJECTIVES: To compare the Seeplex RV15 (test of record) with two newer generation multiplex assays, the Anyplex II RV16 and the xTAG respiratory virus panels. STUDY DESIGN: Two hundred and three retrospective and 36 prospective respiratory samples were tested by all three assays...
June 17, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28633757/the-relationship-between-circulating-mitochondrial-dna-and-inflammatory-cytokines-in-patients-with-major-depression
#17
Yuki Kageyama, Takaoki Kasahara, Masaki Kato, Shiho Sakai, Yasuhiko Deguchi, Munehide Tani, Kenji Kuroda, Kotaro Hattori, Sumiko Yoshida, Yuichi Goto, Toshihiko Kinoshita, Koki Inoue, Tadafumi Kato
BACKGROUND: Although inflammatory cytokines are established biomarkers of mood disorders, their molecular mechanism is not known. We hypothesized that circulating mitochondrial DNA (mtDNA) contributes to inflammation and could be used as biomarkers. We investigated if circulating mtDNA level is associated with inflammatory cytokines and can be used as a biomarker of mood disorders. METHODS: Plasma mtDNA concentration was measured with real-time quantitative PCR targeting two regions of the mtDNA and plasma levels of four cytokines (GM-CSF, IL-2, IL-4, and IL-6) were measured with a multiplex immunoassay method in 109 patients with major depressive disorder (MDD)...
June 6, 2017: Journal of Affective Disorders
https://www.readbyqxmd.com/read/28630585/identification-of-the-plasmodium-species-in-clinical-samples-from-children-residing-in-five-epidemiological-strata-of-malaria-in-cameroon
#18
Tebit Emmanuel Kwenti, Tayong Dizzle Bita Kwenti, Longdoh Anna Njunda, Andreas Latz, Kukwah Anthony Tufon, Theresa Nkuo-Akenji
BACKGROUND: Malaria in Cameroon was previously known to be caused solely by Plasmodium falciparum but today, evidence points to other Plasmodium species including P. vivax, P. ovale and P. malariae. The purpose of this study was to identify the Plasmodium species in clinical samples from children residing in five epidemiological strata of malaria in Cameroon, so as to advise control policies. METHODS: One thousand six hundred nine febrile children (≤15 years) were recruited from five epidemiological strata of malaria including the Sudano-sahelian (SS) strata, the High inland plateau (HIP) strata, the South Cameroonian Equatorial forest (SCEF) strata, the High western plateau (HWP) strata and the Coastal (C) strata...
2017: Tropical Medicine and Health
https://www.readbyqxmd.com/read/28629969/prevalence-and-serotype-distribution-of-listeria-monocytogenes-isolated-from-foods-in-montevideo-uruguay
#19
Valeria Braga, Sylvia Vázquez, Victoria Vico, Valeria Pastorino, María Inés Mota, Marcela Legnani, Felipe Schelotto, Gustavo Lancibidad, Gustavo Varela
The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese) were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment...
June 3, 2017: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/28629316/a-multiplex-primer-design-algorithm-for-target-amplification-of-continuous-genomic-regions
#20
Ahmet Rasit Ozturk, Tolga Can
BACKGROUND: Targeted Next Generation Sequencing (NGS) assays are cost-efficient and reliable alternatives to Sanger sequencing. For sequencing of very large set of genes, the target enrichment approach is suitable. However, for smaller genomic regions, the target amplification method is more efficient than both the target enrichment method and Sanger sequencing. The major difficulty of the target amplification method is the preparation of amplicons, regarding required time, equipment, and labor...
June 19, 2017: BMC Bioinformatics
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