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Molecular and Cellular Probes

Lin Zhang, Dongmei Lin, Haiyan Li, Sen Yu, Junping Bai, Zhiyong Ding, Jiaqiang Wu
Vaccination is an important approach for the control of avian viral diseases. The aim of this study was to evaluate the immune-potentiating effect of oral administration of Inonotus obliquus fermentation products (IOFP) at vaccination in chickens. In total, 120 one-day-old specific-pathogen-free chickens were randomly assigned to six groups: groups 1 to 3 were vaccinated with Newcastle disease virus (NDV) LaSota live vaccine via intranasal and eye-dropped route at seven days of age, and boosted two weeks later...
September 15, 2018: Molecular and Cellular Probes
Aline Aparecida Silva Stella, Larissa Fernanda Simielli Fonseca, Daniele Fernanda Jovino Gimenez, Fernanda Tanamati, André Vieira do Nascimento, Diercles Francisco Cardoso, Humberto Tonhati
This study compared the expression profile of the candidate genes, CSF3 and LPO, by investigating the immune response mechanisms involved in the phenotype of resistance and susceptibility to mastitis of healthy and infected buffaloes. The Granulocyte Colony Stimulating Factor 3 (CSF3) and Lactoperoxidase (LPO) genes expression profiles were determined in 24 milk samples from buffaloes with (N = 12) and without (N = 12) mastitis, using the quantitative real-time PCR (qRT-PCR) technique. CSF3 and LPO expressions were 5...
September 11, 2018: Molecular and Cellular Probes
Jing Chen, Yuanyang Wang, Xiaoqing Liu, Guopei Chen, Xuejian Chen, Jiaping Chen, Zhongdong Liu, Jingwen Gong, Guowu Yang, Quanxue Lan
Streptococcus pyogenes (Group A Streptococcus, GAS) and Streptococcus agalactiae (Group B Streptococcus, GBS) are common pathogens that threaten public health. In this study, a double recombinase polymerase (RPA) amplification assay was developed to rapidly detect these pathogens. Specificity tests revealed that the GAS and GBS strains were positive for speB and SIP genes, respectively. In clinical samples, the double assay performed similarly to the traditional biochemical method. The limits of detection were both ≤100 copies per reaction...
August 29, 2018: Molecular and Cellular Probes
Lei Ma, Feng Cong, Yujun Zhu, Miaoli Wu, Fengjiao Xu, Ren Huang, Robert J Moore, Pengju Guo
Theiler's murine encephalomyelitis virus (TMEV) is one of the most common viral pathogens that circulate widely in captive mouse colonies. A molecular biology detection method would be a useful tool to use in an integrated program to monitor and prevent TMEV infection and transmission. Thus, a reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed to detect TMEV infection. The sensitivity of the RT-RPA assay approached 8 copies per reaction, which is equivalent to the sensitivity of RT-qPCR reactions...
August 26, 2018: Molecular and Cellular Probes
Amir Jahic, Andrea Bock, Franz Duca, Diana Bonderman, Julia Mascherbauer, Reinhard Windhager, Michaela Auer-Grumbach, Christian Beetz
TTR amyloidosis (ATTR) is a fatal condition caused by extracellular deposits of misfolded transthyretin. Patients often present with cardiac disease, but manifestations may also involve other organs including the peripheral nervous system. ATTR is considered familial when heterozygous mutations in the TTR gene are present (ATTRmutant or ATTRm), or acquired when no TTR aberrations are detected (ATTRwildtype or ATTRwt). We hypothesized that TTR copy number variants (CNVs), which would escape the standard diagnostic approaches, contribute to ATTR-related phenotypes, and developed a multiplex ligation-dependent probe amplification-based (MLPA-based), TTR-specific copy number screening tool...
August 21, 2018: Molecular and Cellular Probes
Guimin Zhao, Hongmei Wang, Peili Hou, Xianzhu Xia, Hongbin He
Bovine respirovirus 3 also known as Bovine parainfluenza virus type 3 (BPIV3) is one of the most important viral respiratory agents of both young and adult cattle. Rapid diagnosis could contribute greatly in containing epidemics and thus avoid economic losses. However, the lack of robust isothermal visual method poses difficulty. In this study, a novel isothermal assay for detecting BPIV3 was established. The method includes a lateral flow dipstick (LFD) assay combined with reverse transcription recombinase polymerase amplification (RT-RPA)...
August 20, 2018: Molecular and Cellular Probes
Yana Pestova, Olga Byadovskaya, Alexander Kononov, Alexander Sprygin
In this paper, we report on the development of a real time high-resolution melting (HRM) PCR assay for detection and differentiation among sheep pox virus (SPPV), goat pox virus (GTPV), field isolates and vaccine strains of lumpy skin disease virus (LSDV) based on high-resolution melting curve analysis of their target PCR amplicons. A 111 bp region of LSDV010 ORF, which harbors unique genetic differences for each of these viral species, was selected as the PCR target in this study. During the validation of this assay using DNA from clinical isolates originated from naturally infected animals from the different geographic locations and reference strains, the obtained PCR amplicons demonstrated that the melting temperature picks were specific for each tested viral species, i...
August 10, 2018: Molecular and Cellular Probes
Peres Ramos Badial, Raffaella Bertoni Cavalcanti Teixeira, Diego José Zanzarini Delfiol, Ligia Souza Lima Silveira da Mota, Alexandre Secorun Borges
Overo lethal white foal syndrome (OLWFS) is a genetic disorder caused by a dinucleotide mutation in the endothelin receptor type B (EDNRB) gene leading to the death of affected foals shortly after birth. The use of rapid and reliable genetic testing is imperative for the early diagnosis of the mutation avoiding, therefore, either additional suffering or the production of affected animals. In the present study, we developed and validated a high-resolution melting (HRM) genotyping assay to detect the OLWFS causative mutation, and we also determined the frequency of heterozygotes among American Paint horses in Brazil...
August 8, 2018: Molecular and Cellular Probes
Wei Lu, Lin Wang, Jing Yao, Wen Wang, Yu Chen
C5a is an important pro-inflammatory peptide involved in complement activation, membrane attack complex formation, immune cell chemotaxis, and allergic responses. Osteoarthritis is a disease characterized by degenerative changes in articular cartilage. It has recently been found that inflammatory responses play an important role in the pathogenesis of osteoarthritis and also in rheumatoid arthritis, where dysfunctional synoviocytes are involved. We performed a series of studies to verify our hypothesis that inhibition of C5a would prevent IL-1β-induced alternations in rat synoviocytes...
August 6, 2018: Molecular and Cellular Probes
Reza Shahbazi, Mojtaba Salouti, Bahram Amini, Ahmad Jalilvand, Ebrahim Naderlou, Ali Amini, Arash Shams
Staphylococcus aureus is a gram-positive and opportunistic pathogen that is one of the most common causes of nosocomial infections; therefore, its rapid diagnosis is important and valuable. Today, the use of nanoparticles is expanding due to their unique properties. The purpose of the present study is the determination of S. aureus by a colorimetric method based on gold nanoparticles (AuNPs). Firstly, S. aureus was cultured on both LB media (broth and agar) and their chromosomal DNA was extracted. Afterwards, primers and biosensor were designed based on Protein A sequence data in the gene bank...
July 25, 2018: Molecular and Cellular Probes
Morakot Sroyraya, Sineenart Songkoomkrong, Narin Changklungmoa, Jaruwan Poljaroen, Sawaek Weerakiet, Areepan Sophonsritsuk, Anna Wongkularb, Srithean Lertvikool, Yada Tingthanatikul, Prasert Sobhon
BACKGROUND: Depo-medroxyprogesterone acetate (DMPA) is an injectable progestin contraceptive that provides a highly effective reduction of pelvic pain in women with endometriosis. Despite its wide use to treat pain associated with endometriosis, its precise mechanisms of action remain unclear. The aims of this study were to investigate the differential expressions of estrogen receptors (ERs), and progesterone receptors (PRs) in endometria and ovarian endometrioma cyst walls of women with endometriosis with and without DMPA treatment...
August 2018: Molecular and Cellular Probes
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No abstract text is available yet for this article.
August 2018: Molecular and Cellular Probes
Zili Chen, Yifei Ma, Yaozhen Pan, Haitao Zhu, Chao Yu, Chengyi Sun
Dysregulation of miR-1297 has been detected in various human cancers, and miR-1297 can function as either an oncogene or tumor suppressor. However, the role of miR-1297 in pancreatic adenocarcinoma has not been previously reported. Here, we investigated miR-1297 expression in pancreatic cancer and the role it plays in the development and metastasis of pancreatic adenocarcinoma. In the present study, MiR-1297 and metadherin (MTDH) expression in pancreatic cancer tissue was detected using quantitative real-time PCR (qRT-PCR) and western blot methods...
August 2018: Molecular and Cellular Probes
Ping Guo, Yongxin Yu, Yingjie Pan, Shuling Yan, Yongjie Wang
A pair of nested PCR universal primers (NGIOF and NGIOR) specific for genogroup I (GI) noroviruses was designed based on all GI sequences available in public databases. The primers were evaluated for their specificity, sensitivity and coverage, which demonstrate their reliable performance upon detection of GI noroviruses in oysters.
August 2018: Molecular and Cellular Probes
Marco Ligozzi, Liliana Galia, Maria Carelli, Pier Paolo Piccaluga, Erica Diani, Davide Gibellini
In this study, we describe a duplex real-time PCR assay for the simultaneous detection of KIPyV and WUPyV polyomaviruses based on TaqMan probes. This assay detected 500 copies/mL both for KIPyV and WUPyV in 100% of tested positive samples. We assessed this technique on 482 nasopharyngeal aspirate specimens from hospitalized pediatric patients with respiratory symptoms, previously analyzed with commercial multiplex assay for 16 major respiratory viruses. Our assay detected KIPyV genome in 15 out of 482 samples (3...
August 2018: Molecular and Cellular Probes
Zhihua Ju, Jinming Huang, Qiang Jiang, Changfa Wang, Xiuge Wang, Shuhong Zhao
Bovine mastitis is an inflammation response of the mammary gland tissues caused mainly by pathogenic bacteria in cows. Previous studies showed that bta-miR-15a and bta-miR-16a modulate immunity and inflammation responses. In this study, we investigated the expression pattern and tissue localization of bta-miR-15a and bta-miR-16a. The expression levels of bta-miR-15a and bta-miR-16a were significantly upregulated in mammary tissues and blood neutrophils of mastitis-infected cows, compared with those of healthy cows (P < 0...
August 2018: Molecular and Cellular Probes
Feng Lin, Li Liu, Gui-Jie Hao, Peng-Chen Sheng, Zheng Cao, Yang Zhou, Peng Lv, Ting Xu, Jinyu Shen, KePing Chen
White tail disease (WTD), a major disease prevailing in the larval stage of Macrobrachium rosenbergii, caused by Macrobrachium rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), led to the economic loss of shrimp industry in China. In order to establish a convenient, sensitive and selective molecular diagnostic method to detect MrNV and XSV for the Chinese shrimp (MrNV/XSV-chin), a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay combined with a lateral flow dipstick (LFD) method were developed...
August 2018: Molecular and Cellular Probes
Zhiqing Huang, Jianwei Zheng, Chunmei Shi, Qiang Chen
Coupling propidium monoazide (PMA) with quantitative PCR (PMA-qPCR) has been successfully applied to specific detection and quantification of viable cells in various samples. The optimal PMA treatment condition is usually determined through qPCR. However, it is a tedious, time consuming and costly process including DNA extraction and qPCR. To overcome this problem, a flow cytometry-based (FCM-based) method was first proposed in this study to replace qPCR for screening of the optimal PMA treatment condition for Helicobacter pylori, since the pure culture treated with PMA was actually a single cell suspension with fluorescent dye...
August 2018: Molecular and Cellular Probes
Ronald J A Wanders
Peroxisomes catalyze a number of essential metabolic functions of which fatty acid alpha- and beta-oxidation, ether phospholipid biosynthesis, glyoxylate detoxification and bile acid synthesis are the most important. The key role of peroxisomes in humans is exemplified by the existence of a group of peroxisomal disorders, caused by mutations in > 30 different genes which code for proteins with a role in either peroxisome biogenesis or one of the metabolic pathways in peroxisomes. Technological advances in laboratory methods at the metabolite-, enzyme-, and molecular level have not only allowed the identification of new peroxisomal disorders but also new phenotypes associated with already identified genetic defects thus extending the clinical spectrum...
August 2018: Molecular and Cellular Probes
Ruthellen H Anderson, Kevin R Francis
Rare diseases, in totality, affect a significant proportion of the population and represent an unmet medical need facing the scientific community. However, the treatment of individuals affected by rare diseases is hampered by poorly understood mechanisms preventing the development of viable therapeutics. The discovery and application of cellular reprogramming to create novel induced pluripotent stem cell models of rare diseases has revolutionized the rare disease community. Through developmental and functional analysis of differentiated cell types, these stem cell models carrying patient-specific mutations have become an invaluable tool for rare disease research...
August 2018: Molecular and Cellular Probes
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