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Molecular and Cellular Probes

Liang Wang, Guo-Fu Chen, Chun-Yun Zhang, Yuan-Yuan Wang, Rui Sun
Frequent outbreaks of toxic algal blooms devastate marine ecosystems, marine fisheries, and public health. Monitoring toxic algae is crucial to reduce losses caused by imminent algal blooms. However, traditional detection techniques relying on morphological examination are tedious and time-consuming. Therefore, the development of convenient strategies to detect toxin-producing microalgae is necessary. In this study, a novel method for the rapid, sensitive detection of Amphidinium carterae by loop-mediated isothermal amplification (LAMP) combined with a chromatographic lateral-flow dipstick (LFD) was established...
November 9, 2018: Molecular and Cellular Probes
Jingjiao Ma, Ye Tian, Min Zhang, Yujie Li, Weili Wang, Fulin Tian, Yuqiang Chen, Yaxian Yan, Jianhe Sun
Budgerigar fledgling disease virus (BFDV) infection causes sudden death, abdominal distention, and feather abnormality in psittacine birds. In this study, we developed a TaqMan Real-time PCR assay to detect BFDV by targeting a conserved region in VP1 gene. The detection limit of the assay was 30 DNA gene copies, 1000 times more sensitive than conventional PCR. The coefficients of variation were less than 1.09% in either intra- or inter-assays, indicating high reproducibility. By using this method, the prevalence of BFDV in China was evaluated...
November 8, 2018: Molecular and Cellular Probes
Ning Sun, Weiping Wang, Jie Wang, Xinyue Yao, Fangfang Chen, Xiaojun Li, Yi Yinglei, Bo Chen
Three reverse transcription recombinase polymerase amplification assays with lateral flow dipsticks (RT-RPA-LFD) were developed for identification of the matrix and hemagglutinin (HA) genes to detect influenza A virus and distinguish subtypes H1 and H3. Assessment of the assays' specificity showed that there was no cross-reactivity with other targets. Their limits of detection were 123.6 copies per reaction for the matrix gene, 677.1 copies per reaction for the H1 HA gene, and 112.2 copies/reaction for the H3 HA gene...
October 27, 2018: Molecular and Cellular Probes
Jae-Ok Kim, Jong-Oh Kim, Soo-Jin Kim, Wi-Sik Kim, Myung-Joo Oh
In situ hybridization (ISH) of genomic segments using RNA-RNA hybrid for nervous necrosis virus (NNV) detection has not been reported yet. The objective of this study was to develop RNA-ISH using RNA probes for the detection of NNV in infects SSN-1 cells or sevenband grouper Hyporthodus septemfasciatus. Two viral RNA segments viz., RNA1 and RNA2 were synthesized by in vitro transcription and labeled with fluorescein UTP and dignoxigenin dUTP, respectively. These labeled RNA probes specifically detected NNV in infected SSN-1 cells...
October 18, 2018: Molecular and Cellular Probes
Xueyao Ma, Dawei Wei, Gong Cheng, Shijun Li, Li Wang, Yaning Wang, Xiaoyu Wang, Song Zhang, Hongbao Wang, Linsen Zan
Deposition of intramuscular fat (IMF) is one of the most important traits for the evaluation of beef carcass quality grade. MicroRNA (miRNA) is an endogenous non-coding RNA that can play a role in the post-transcriptional regulation of mammalian preadipocyte differentiation. Previously, we identified that bta-miR-130a regulates milk fat biosynthesis by targeting PPARG mRNA. However, the role of miR-130 in the regulation of bovine adipocyte differentiation remains unknown. In this study, we found that overexpression of bta-miR-130a/b led to significantly decreased cellular triacylglycerol (TAG) levels during adipogenesis process as well as reduced lipid droplet formation...
October 15, 2018: Molecular and Cellular Probes
Myung-Jin Mun, Joon-Yong Bae, Jin Hyuck Kim, Soo Bok Kim, Ilseob Lee, Jin Il Kim, Mee Sook Park, Man-Seong Park, Yong Suk Nam
Previous studies reported that severity of dengue is associated with multiple factors, including secondary infection, age, viral load and infecting serotype and genotype. In addition, other studies have reported that a dengue virus-2 (DENV-2) infection is associated with a prognosis of more severe clinical manifestations than DENV-1 and DENV-4 infections. For these reasons, the ability to identify the DENV serotypes is critical for optimal patient diagnosis and epidemiological studies. In this study, we developed a TaqMan probe-based, one-step real-time reverse transcriptase-polymerase chain reaction (RT-PCR) system for detection and serotyping DENV...
October 3, 2018: Molecular and Cellular Probes
Muhammet Uslupehlivan, Remziye Deveci, Cemal Ün
The prion protein is a membrane-bound glycoprotein which consists mainly α-helix structure. In contrast, the infectious prion protein shows the beta-sheet structure. The prion-associated diseases are all lethal neurodegenerative abnormalities, called transmissible spongiform encephalopathies. Scrapie is the most common type of these illnesses affecting sheep, goats, and moufflon. The VRQ, AHQ, ARR and N146S polymorphisms in the sheep prion gene have been found to be associated with resistance to scrapie disease...
September 24, 2018: Molecular and Cellular Probes
Shao Wang, Shifeng Xiao, Xiaoxia Cheng, Shaoying Chen, Xiaoli Zhu, Fengqiang Lin, Shilong Chen
To obtain a deletion mutant of Muscovy duck-origin goose parvovirus (MDGPV) and to analyze its biological characteristics, the pMDGPVPT plasmid, which contains a full-length DNA infectious clone of the MDGPV PT strain, was used in this study as the template. The E-box at nt 315 of the left inverted terminal repeat sequence (L-ITR) was deleted by overlap extension PCR to obtain the infectious recombinant plasmid p-PTΔE315 . The p-PTΔE315 plasmid was transfected into 9-day-old non-immune Muscovy duck embryos via the yolk sac and the rescued deletion mutant virus r-PTΔE315 was generated...
September 18, 2018: Molecular and Cellular Probes
Yaser Ghazi, Fatemeh Haddadi, Hossein Kamaladini
The conventional techniques of PCR, Southern blot, northern blot, in situ hybridization, and RNase protection assay have long been used to investigate transformation and expression of genes, but most of them are time-consuming and have relatively low sensitivity. In recent years, applying biosensors for molecular identification of biomolecules has been expanding significantly. Hence in this study, Zabol melon was used as a model plant to introduce new DNA and RNA-based biosensors for confirming gene transformation and expression...
October 2018: 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...
October 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...
October 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...
October 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...
October 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...
October 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)...
October 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...
October 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...
October 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...
October 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...
October 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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