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Molecular and Cellular Probes

Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Wasim Shehzad, Peter C Thomson
For rapid and simultaneous detection of nine bovine mastitic pathogens, a sensitive and specific multiplex PCR assay was developed. The assay was standardized using reference strains and validated on mastitic milk cultures which were identified to species level based on 16S rRNA sequencing. Multiplex PCR assay also efficiently detected the target bacterial strains directly from milk. Detection limit of the assay was up to 50 pg for DNA isolated from pure cultures and 10(4) CFU/ml for spiked milk samples. As estimated by latent class analysis, the assay was sensitive up to 88% and specific up to 98% for targeted mastitic pathogens, compared with the bacterial culture method and the 16S rRNA sequence analysis...
March 20, 2017: Molecular and Cellular Probes
Chong Cao, Xin Luo, Xiaowei Ji, Yao Wang, Yuan Zhang, Pengtao Zhang, Liangjun Zhong
OBJECTIVE: The present study aimed to explore the possible effects of osteopontin (OPN) in the proliferation of rat aortic smooth muscle cells (RASMCs) stimulated by gingipains. METHODS: The proliferation of RASMCs in response to active gingipains treatment was evaluated by CCK-8 assay. OPN siRNA was designed, constructed and transfected into RASMCs at different concentrations. The cell cycle of RASMCs was analyzed by flow cytometry. OPN, α-SMA and calponin expression were examined by real-time PCR and western blot analysis...
March 13, 2017: Molecular and Cellular Probes
Alais Maria Dall Agnol, Rodrigo Alejandro Arellano Otonel, Raquel Arruda Leme, Amauri Alcindo Alfieri, Alice Fernandes Alfieri
This study describes a sensitive (1.3 × 10(1) genomic copies/μL) and specific TaqMan-based qRT-PCR assay able to detect and quantify SVA RNA in porcine biological samples. The technique represents an efficient tool for the virus diagnosis and assessment of SVA load in tissues of infected animals and for epidemiological studies.
March 4, 2017: Molecular and Cellular Probes
Maria Pia Leone, Pietro Palumbo, Rocco Ortore, Stefano Castellana, Orazio Palumbo, Salvatore Melchionda, Teresa Palladino, Raffaella Stallone, Tommaso Mazza, Roberto Cocchi, Massimo Carella
The paper describes a putative digenic form of deafness in two siblings affected by non-syndromic hereditary hearing loss, detected by a Targeted resequencing approach. Given that a previous paper suggested TMPRSS3 and GJB2 genes as responsible for a digenic form of hearing loss, our data support and reinforce this hypothesis.
March 3, 2017: Molecular and Cellular Probes
Maira N Meggiolaro, Anna Ly, Benjamin Rysnik-Steck, Carolina Silva, Joshua Zhang, Damien P Higgins, Gary Muscatello, Jacqueline M Norris, Mark Krockenberger, Jan Šlapeta
Canine parvovirus (CPV-2) remains an important cause of devastating enteritis in young dogs. It can be successfully prevented with live attenuated CPV-2 vaccines when given at the appropriate age and in the absence of maternal antibody interference. Rapid diagnosis of parvoviral enteritis in young dogs is essential to ensuring suitable barrier nursing protocols within veterinary hospitals. The current diagnostic trend is to use multiplexed PCR panels to detect an array of pathogens commonly responsible for diarrhea in dogs...
February 27, 2017: Molecular and Cellular Probes
Adán Valenzuela-Castillo, Fernando Mendoza-Cano, Tania Enríquez-Espinosa, José Manuel Grijalva-Chon, Arturo Sánchez-Paz
The decapod Penstyldensovirus 1 (PstDV-1) represents one of the most serious threats for penaeid shrimp farming. Studies aimed at defining relevant molecular effects of this virus over its host are imperative in the attempt to increase our understanding of its pathogenesis. Unfortunately, few studies have focused on the definition of the expression profile of reference genes in shrimp challenged with a pathogen. As a result, there are no studies on the selection of reference genes for the normalization of target gene expression changes yielding reliable data of the effects following PstDV-1 infection in shrimp...
February 20, 2017: Molecular and Cellular Probes
Ahmed S Abdel-Moneim, Gaber M Shehab, Adnan A Alsulaimani, Mater I R Al-Malky
The human respiratory syncytial virus is a common respiratory pathogen in children. Improved diagnosis of the virus is dependent on the development of tools for the rapid detection and estimation of the viral loads. In the current study, RT-qPCR using TaqMan hydrolysis probe based on the F gene detection was developed to identify and quantify hRSV in clinical samples. The assay was validated by comparing the results with a commercially available RT-qPCR kit. The newly developed assay was sensitive in detecting hRSV positive samples (59/126) which were equivalent to those detected by the commercial kit (57/126) with a detection limit of 1 × 10(2) copies/mL...
February 20, 2017: Molecular and Cellular Probes
Ruslan Kalendar, Alexander Belyayev, Tatiana Zachepilo, Alexander Vaido, Dmitry Maidanyuk, Alan H Schulman, Natalia Dyuzhikova
We evaluated copy number variation (CNV) for four genes in rat strains differing in nervous system excitability. rpl13a copy number is significantly reduced in hippocampus and bone marrow in rats with a high excitability threshold and stress. The observed phenomenon may be associated with a role for rpl13a in lipid metabolism.
February 10, 2017: Molecular and Cellular Probes
Jingfang Chen, Rusheng Zhang, Xinhua Ou, Dong Yao, Zheng Huang, Linzhi Li, Biancheng Sun
A TaqMan based duplex one-step real time RT-PCR (rRT-PCR) assay was developed for the rapid detection of Coxsackievirus A10 (CV-A10) and other enterovirus (EVs) in clinical samples. The assay was fully evaluated and found to be specific and sensitive. When applied in 115 clinical samples, a 100% diagnostic sensitivity in CV-A10 detection and 97.4% diagnostic sensitivity in other EVs were found.
February 8, 2017: Molecular and Cellular Probes
Xinrong Zhou, Tiansheng Zhang, Deping Song, Tao Huang, Qi Peng, Yanjun Chen, Anqi Li, Fanfan Zhang, Qiong Wu, Yu Ye, Yuxin Tang
Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal disease, resulting in substantial economic losses to the swine industry worldwide. In this study, three assays, namely a conventional reverse transcription-polymerase chain reaction (RT-PCR), a SYBR Green I real-time RT-PCR and a TaqMan real-time RT-PCR targeting the highly conserved M gene of PEDV, were developed and evaluated. Then, the analytical specificity, sensitivity and reproducibility of these assays were determined and compared...
February 7, 2017: Molecular and Cellular Probes
Liyuan Liu, Luzhi Jiang, Yongxin Yu, Xiaoming Xia, Yingjie Pan, Shuling Yan, Yongjie Wang
A rapid and sensitive AHPND-RPA assay was developed for the specific detection of the AHPND-causing Vibrio owensii. The AHPND-RPA detected as few as 2 copies per reaction in 9.02 ± 0.66 min at 39 °C, and showed 100% positive predictive value and negative predictive value for AHPND diagnosis.
February 5, 2017: Molecular and Cellular Probes
Ghazaleh Farmanfarmaei, Mansour Kargarpour Kamakoli, Hamid Reza Sadegh, Morteza Masoumi, Farid Abdolrahimi, Abolfazl Fateh, Nayereh Ebrahimzadeh, Gholamreza Javadi, Fatemeh Rahimi Jamnani, Farzam Vaziri, Seyed Davar Siadat
The application of MIRU-VNTR has unveiled that infection by Mycobacterium tuberculosis can be polyclonal. Our comparative study demonstrated that based on the studied samples (clinical specimen or culture) detection of polyclonal M. tuberculosis infection can be significantly different.
January 24, 2017: Molecular and Cellular Probes
Hans-Peter Fuehrer, Sara Savić
No abstract text is available yet for this article.
February 2017: Molecular and Cellular Probes
Sally J Cutler, Eva Ruzic-Sabljic, Aleksandar Potkonjak
Lyme borreliosis (or Lyme disease) has become a virtual household term to the exclusion of other forgotten, emerging or re-emerging borreliae. We review current knowledge regarding these other borreliae, exploring their ecology, epidemiology and pathological potential, for example, for the newly described B. mayonii. These bacteria range from tick-borne, relapsing fever-inducing strains detected in some soft ticks, such as B. mvumii, to those from bat ticks resembling B. turicatae. Some of these emerging pathogens remain unnamed, such as the borrelial strains found in South African penguins and some African cattle ticks...
February 2017: Molecular and Cellular Probes
Jacinto Gomes, Marcos Santos, Ana Amaro, Isabel Pereira da Fonseca, Gabriela Santos-Gomes, João Inácio
A loop-mediated isothermal amplification (LAMP) assay was evaluated for the detection of Theileria annulata infection in cattle. The results were compared with a real-time PCR used for the quantification of T. annulata parasitaemia. One hundred bovine blood samples from 16 cattle farms were tested with LAMP and real-time PCR, with T. annulata DNA being detected in 66% and 67% of the samples, respectively. The results showed that the LAMP assay detects a parasitaemia as low as 0.00025%, indicating a high analytical sensitivity of LAMP for clinical diagnosis of bovine theileriosis...
December 22, 2016: Molecular and Cellular Probes
Hedayatollah Shirzad, Fariba Esmaeili, Shabnam Bakhshalizadeh, Marzieh Ebrahimie, Esmaeil Ebrahimie
Murine P19 embryonal carcinoma (EC) cells are convenient to differentiate into all germ layer derivatives. One of the advantages of P19 cells is that the exogenous DNA can be easily inserted into them. Here, at the first part of this study, we generated stable GFP-expressing P19 cells (P19-GFP(+)). FACS and western-blot analysis confirmed stable expression of GFP in the cells. We previously demonstrated the efficient induction of neuronal differentiation from mouse ES and EC cells by application of a neuroprotective drug, selegiline In the second part of this study selegiline was used to induce differentiation of P19-GFP(+) into stable GFP-expressing neuron-like cells...
December 21, 2016: Molecular and Cellular Probes
Miguel Angel Ayala-Valdovinos, Jorge Galindo-García, David Sánchez-Chiprés, Theodor Duifhuis-Rivera
Hydrocephalus in Friesian horses is an autosomal recessive hereditary disease that can result in an abortion, a stillbirth, or euthanization of a newborn foal. Here, the hydrocephalus-associated c.1423C > T mutation in B3GALNT2 gene was detected with PCR-RFLP and PCR-PIRA methods for horse genotyping. A preliminary genotyping survey was performed on 83 randomly selected Friesian stallion horses to determine the current allele frequency in Mexico. The frequency of the mutant T allele was 9.6%.
December 21, 2016: Molecular and Cellular Probes
Gaurava K Rai, Rubina Lawrence
Making specific oligonucleotide probes for diagnosis and differentiation of viruses has always been a challenging task due to unavailability of defined viral sequence databases, probe designing softwares and higher degree of sequence homology among the types and subtypes of viruses. We have provided here, a streamlined procedure to design microarray probes involving various computational tools which is capable of yielding highly specific as well as conserved oligonucleotides to facilitate unbiased diagnosis of viruses...
December 15, 2016: Molecular and Cellular Probes
Rebecca Hinz, Norbert G Schwarz, Andreas Hahn, Hagen Frickmann
Schistosomiasis is a common disease in endemic areas of Sub-Saharan Africa, South America and Asia. It is rare in Europe, mainly imported from endemic countries due to travelling or human migration. Available methods for the diagnosis of schistosomiasis comprise microscopic, molecular and serological approaches, with the latter detecting antigens or antibodies associated with Schistosoma spp. infection. The serological approach is a valuable screening tool in low-endemicity settings and for travel medicine, though the interpretation of any diagnostic results requires knowledge of test characteristics and a patient's history...
December 13, 2016: Molecular and Cellular Probes
Linsheng Gui, Jieyun Hong, Sayed Haidar Abbas Raza, Linsen Zan
Sirtuin 3 (SIRT3) is a mitochondrial nicotinamide adenine dinucleotide (NAD)-dependent deacetylase. It has crucial roles in regulating the respiratory chain, in adenosine triphosphate (ATP) production, and in both the citric acid and urea cycles. The aim of this study was to investigate whether SIRT3 could be used as a candidate gene in the breeding of cattle. Expression analysis by quantitative real-time polymerase chain reactions (qPCR) indicated that expression levels of SIRT3 were highest in the kidney, rumen, liver, omasum and muscle...
December 12, 2016: Molecular and Cellular Probes
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