scholarly journals Genotipos de VPH y cambios citológicos cervico-uterino en pacientes de una consulta ginecológica privada del Estado Carabobo, Venezuela. Marzo-octubre de 2017

2020 ◽  
Vol 26 (1) ◽  
Author(s):  
Monica Alejandra Sequera ◽  
Andrea Matamoros ◽  
Maria Jose Mendoza León
Keyword(s):  
Multiple Pcr

Introducción: La infección genital por el Virus de Papiloma Humano (VPH) se ha asociado con el cáncer cérvicouterino (CCE) al provocar la aparición de lesiones precursoras de cáncer en la zona de transformación de la unión escamo-columnar del cuello uterino. Existen más de 100 tipos de VPH, clasificados en bajo riesgo oncogénico (VPH-BR) y alto riesgo oncogénico (VPH-AR). Estudios reportan la infección por genotipos de alto riesgo en el 100% de los CCE. En Venezuela, el 67,7% de los CCE, se relacionan con el genotipo de VPH-AR 16. Objetivo: Detectar la presencia de VPH en pacientes con cambios citológicos cervicouterino. Metodología: Se incluyeron 49 pacientes que presentaban cambios citológicos, se tomaron las muestras de la región endocervical y exocervical para la detección y genotipificación del virus mediante la técnica de Multiple PCR. Resultados: Las alteraciones citológicas presentes fueron Células Escamosas Atípicas (69,4%), Células Glandulares Atípicas (4,1%), Lesión Escamosa Intraepitelial de Bajo Grado (16,3%), y Lesión Escamosa Intraepitelial de Alto Grado (10,2%). La detección molecular demostró que 16,3% presentaba VPH, 62,5% correspondían a VPH-AR, 25% a VPH-BR, 12,5% al genotipo 16 y no se detectó el genotipo 18. Se reportó un solo caso de coinfección. Conclusiones: A diferencia de otros estudios, no se encontró una relación estadísticamente significativa entre la presencia del virus y la aparición de cambios citológicos cervicouterino en esta población. No obstante, se detectaron genotipos de alto riesgo oncogénico, lo que puede traducirse en una mayor incidencia de cáncer cervicouterino a futuro.  

A Multiplex PCR Assay Mediated by Universal Primers for the Detection of Adulterated Meat in Mutton

2019 ◽  
Vol 82 (2) ◽  
pp. 325-330 ◽  
Author(s):  
WANWAN LIU ◽  
XIAONAN WANG ◽  
JING TAO ◽  
BANGSHENG XI ◽  
MAN XUE ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Detection System ◽  
Meat Products ◽  
Pcr Primers ◽  
Rapid Identification ◽  
Food Samples ◽  
Detection Sensitivity ◽  
Universal Primers ◽  
Multiplex Pcr Assay ◽  
Multiple Pcr

ABSTRACT This study aimed to establish a multiplex PCR detection system mediated by “universal primers,” which would be able to determine whether mutton meat contained nonmutton ingredients from rats, foxes, and ducks. Based on the sequence variation of specific mitochondrial genes, nine different multiplex PCR primers were designed, and four kinds of meat products were rapidly identified by electrophoresis using an optimized multiplex PCR system based on the molecular weight differences of the amplified products. Multiplex PCR applications optimized for meat food source from food samples for testing was used to verify the accuracy of the identification method. The results showed that the primers in multiple PCR system mediated by universal primers could be used for the rapid identification of rat, fox, duck, and sheep meat in mutton products, and the detection sensitivity could reach 0.05 ng/μL. The identification of food samples validated the practical value of this method. Therefore, a multiplex PCR system mediated by universal primers was established, which can be used to quickly identify the origin of animal ingredients from rats, foxes, and ducks in mutton products.

scholarly journals Utilisation d’un test multiple basé sur la réaction de polymérase en chaîne pour des enquêtes épidémiologiques sur les hémoparasites des bovins au Mexique

1993 ◽  
Vol 46 (1-2) ◽  
pp. 71-75
Author(s):  
Julio Vicente Figueroa ◽  
J.A. Alvarez ◽  
J.A. Ramos ◽  
C.A. Vega ◽  
G.M. Buening
Keyword(s):  
Anaplasma Marginale ◽  
Babesia Bigemina ◽  
Dot Blot ◽  
Multiple Pcr

Une étude a été effectuée sur la possibilité d'appliquer la technique de la réaction de polymérase en chaîne (PCR) pour la détection simultanée des hémoparasites bovins Babesia bigemina, B. bovis et Anaplasma marginale. Des échantillons de sang de bovins ont été récoltés dans des ranches d'une zone endémique identifiée au préalable dans la péninsule de Yucatan au Mexique, et ont été préparés pour analyse par PCR. Ils ont été soumis à l'amplification de l'ADN dans un tube à réaction contenant des amorces d'oligonucléotides spécifiques pour l'ADN de chaque espèce d'hémoparasite. Les produits de la PCR ont été détectés par hybridation en Dot-Blot de l'acide nucléique utilisant des sondes d'ADN non-radioactives, spécifiques, marquées à la digoxigénine par PCR. 420 échantillons analysés par le test multiple PCR-sonde ADN ont montré des taux de prévalence de 66,7 %, 60,1 et 59,6 % pour B. bigemina, B. bovis et A. marginale, respectivement. L'analyse multiple par PCR a montré que des animaux ayant des infections simples, doubles ou triples pouvaient être détectés par les sondes d'ADN spécifiques. La procédure est proposée comme un outil de valeur pour l'analyse épidémiologique dans les régions où ces espèces d'hémoparasites infectent les bovins simultanément.

scholarly journals Genetic Characteristics and Multiple-PCR Development for Capsular Identification of Specific Serotypes of Campylobacter jejuni

PLoS ONE ◽  
2016 ◽  
Vol 11 (10) ◽  
pp. e0165159 ◽  
Author(s):  
Hao Liang ◽  
Aiyu Zhang ◽  
Yixin Gu ◽  
Yuanhai You ◽  
Jianzhong Zhang ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Genetic Characteristics ◽  
Multiple Pcr

Reversible thermo-pneumatic valves on centrifugal microfluidic platforms

Lab on a Chip ◽  
2015 ◽  
Vol 15 (16) ◽  
pp. 3358-3369 ◽  
Author(s):  
Mohammad Mahdi Aeinehvand ◽  
Fatimah Ibrahim ◽  
Sulaiman Wadi Harun ◽  
Amin Kazemzadeh ◽  
Hussin A. Rothan ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Thermal Energy ◽  
Microfluidic Platforms ◽  
Pneumatic Valves ◽  
Multiple Pcr

Reversible thermo-pneumatic valves (RTPVs) manipulate thermal energy to reversibly block or open microchannels. The valves prevent the evaporation of reagents during thermocycling periods. The cartridge is used for sequential aliquoting to prepare multiple PCR reaction mixtures for the detection of the Dengue virus.

scholarly journals High-Frequency Genetic Recombination and Reactivation of Orthopoxviruses from DNA Fragments Transfected into Leporipoxvirus-Infected Cells

2003 ◽  
Vol 77 (13) ◽  
pp. 7281-7290 ◽  
Author(s):  
Xiao-Dan Yao ◽  
David H. Evans
Keyword(s):  
Vaccinia Virus ◽  
Genetic Recombination ◽  
Virus Genome ◽  
Dna Fragments ◽  
Gene Encoding ◽  
Plaque Purification ◽  
Infected Cells ◽  
Clone And Express ◽  
Recombination Reactions ◽  
Multiple Pcr

ABSTRACT Poxvirus DNA is not infectious because establishing an infection requires the activities of enzymes packaged in the virion. This barrier can be overcome by transfecting virus DNA into cells previously infected with another poxvirus, since the resident virus can provide the trans-acting systems needed to reactivate transfected DNA. In this study we show that cells infected with a leporipoxvirus, Shope fibroma virus (SFV), can reactivate vaccinia virus DNA. Similar heterologous packaging systems which used fowlpox-infected cells to reactivate vaccinia virus have been described, but SFV-infected cells promoted a far more efficient reaction that can produce virus titers exceeding 106 PFU/μg of transfected DNA. SFV-promoted reactions also exploit the hyperrecombinogenic systems previously characterized in SFV-infected cells, and these coupled recombination and reactivation reactions could be used to delete nonessential regions of the vaccinia virus genome and to reconstruct vaccinia virus from overlapping DNA fragments. SFV-catalyzed recombination reactions need only two 18- to 20-bp homologies to target PCR amplicons to restriction enzyme-cut vaccinia virus vectors, and this reaction feature was used to rapidly clone and express a gene encoding fluorescent green protein without the need for plaque purification or selectable markers. The ability of SFV-infected cells to reactivate fragments of vaccinia virus was ultimately limited by the number of recombinational exchanges required and one cannot reconstruct vaccinia virus from multiple PCR fragments spanning essential portions of the genome. These observations suggest that recombination is an integral part of poxvirus reactivation reactions and provide a useful new technique for altering the structure of poxvirus genomes.

Integration of Multiple PCR Amplifications and DNA Mutation Analyses by Using Oligonucleotide Microchip

2001 ◽  
Vol 292 (1) ◽  
pp. 155-160 ◽  
Author(s):  
Sergei V Tillib ◽  
Boris N Strizhkov ◽  
Andrei D Mirzabekov
Keyword(s):  
Dna Mutation ◽  
Multiple Pcr ◽  
Mutation Analyses

scholarly journals Multiple PCR for Detection of Genes Encoding Antibiotic Resistance in Staphylococcus aureus

2017 ◽  
Vol 6 (5) ◽  
pp. 1052-1056
Author(s):  
S. Wilfred Ruban ◽  
P. Raja ◽  
Nithya Quintiol ◽  
Govind Vemala ◽  
K. Porteen
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Genes Encoding ◽  
Multiple Pcr

scholarly journals Prevalence of Mutations in the GJB2, SLC26A4, GJB3, and MT-RNR1 Genes in 103 Children with Sensorineural Hearing Loss in Shaoxing, China

2018 ◽  
Vol 97 (6) ◽  
pp. E33-E38 ◽  
Author(s):  
Hong Yu ◽  
Dan Liu ◽  
Jingqun Yang ◽  
Zhiqiang Wu
Keyword(s):  
Hearing Loss ◽  
Sensorineural Hearing Loss ◽  
Genetic Screening ◽  
Mutant Allele ◽  
Hot Spot ◽  
Pathogenic Mutation ◽  
Sensorineural Hearing ◽  
Common Cause ◽  
Vestibular Aqueduct ◽  
Multiple Pcr

Mutations in the GJB2, SLC26A4, GJB3, and MT-RNR1 genes are known to be a common cause of hearing loss. However, the frequency of hot-spot mutations and genotype-phenotype correlations in patients with sensorineural hearing loss (SNHL) has been less frequently reported. We conducted a study of 103 children—56 boys and 47 girls, aged 5 months to 9 years (mean: 4.1 yr)—with SNHL who underwent genetic screening for 20 hot-spot mutations of the GJB2, SLC26A4, GJB3, and MT-RNR1 genes. Mutations were detected by multiple-PCR-based MALDI-TOF MS assay. At least one mutated allele was detected in 48 patients (46.6%), and 30 patients (29.1%) carried pathogenic mutations. Among all the detected mutations, the most common were GJB2 c.235delC and SLC26A4 c.919-2A>G, with allele frequencies of 23.8 and 6.8%, respectively. At least one mutant allele of SLC26A4 was detected in the 13 patients who had an enlarged vestibular aqueduct (EVA). Almost half of the children with SNHL carried a common deafness-related mutation, and nearly one-third carried a pathogenic mutation. The mutations in SLC26A4 were prevalent and correlated strongly with EVA.

scholarly journals Recovery and Detection of Escherichia coli O157:H7 in Surface Water, Using Ultrafiltration and Real-Time PCR

2009 ◽  
Vol 75 (11) ◽  
pp. 3593-3597 ◽  
Author(s):  
Bonnie Mull ◽  
Vincent R. Hill
Keyword(s):  
Escherichia Coli ◽  
Surface Water ◽  
Real Time ◽  
Real Time Pcr ◽  
Membrane Filtration ◽  
Analytical Techniques ◽  
Immunomagnetic Separation ◽  
Treated Wastewater ◽  
Sample Collection ◽  
Multiple Pcr

ABSTRACT Enterohemorrhagic Escherichia coli O157:H7 (EHEC O157:H7) outbreaks have revealed the need for improved analytical techniques for environmental samples. Ultrafiltration (UF) is increasingly recognized as an effective procedure for concentrating and recovering microbes from large volumes of water and treated wastewater. This study describes the application of hollow-fiber UF as the primary step for concentrating EHEC O157:H7 seeded into 40-liter samples of surface water, followed by an established culture/immunomagnetic-separation (IMS) method and a suite of real-time PCR assays. Three TaqMan assays were used to detect the stx1, stx2, and rfbE gene targets. The results from this study indicate that approximately 50 EHEC O157:H7 cells can be consistently recovered from a 40-liter surface water sample and detected by culture and real-time PCR. Centrifugation was investigated and shown to be a viable alternative to membrane filtration in the secondary culture/IMS step when water quality limits the volume of water that can be processed by a filter. Using multiple PCR assay sets to detect rfbE, stx1, and stx2 genes allowed for specific detection of EHEC O157:H7 from strains that do not possess all three genes. The reported sample collection and analysis procedure should be a sensitive and effective tool for detecting EHEC O157:H7 in response to outbreaks of disease associated with contaminated water.

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