Displaying publications 101 - 120 of 1867 in total

Abstract:
Sort:
  1. Tan LL, Ahmed SA, Ng SK, Citartan M, Raabe CA, Rozhdestvensky TS, et al.
    Food Chem, 2020 Mar 30;309:125654.
    PMID: 31678669 DOI: 10.1016/j.foodchem.2019.125654
    A specialized DNA extraction method and a SYBR Green quantitative polymerase chain reaction (SyG-qPCR) assay were combined to generate a ready-to-use kit for rapid detection of porcine admixtures in processed meat products. Our qPCR assay utilized repetitive LINE-1 elements specific to the genome of Sus scrofa domesticus (pig) as a target and incorporated internal controls. We improved the genomic DNA extraction method, and reduced extraction times to the minimum. The method was validated for specificity, sensitivity (0.001% w/w) and robustness, and values were compared with those of a commercially available kit. We also tested our method using 121 processed food products and consistently detected amplification only in samples containing pork. Due to its efficiency and cost-effectiveness, our method represents a valuable new method for detecting food adulteration with pork that is superior to existing quality control approaches.
    Matched MeSH terms: Real-Time Polymerase Chain Reaction/instrumentation; Real-Time Polymerase Chain Reaction/methods*
  2. Selvarajah GT, Bonestroo FAS, Timmermans Sprang EPM, Kirpensteijn J, Mol JA
    BMC Vet Res, 2017 Nov 25;13(1):354.
    PMID: 29178874 DOI: 10.1186/s12917-017-1281-3
    BACKGROUND: Quantitative PCR (qPCR) is a common method for quantifying mRNA expression. Given the heterogeneity present in tumor tissues, it is crucial to normalize target mRNA expression data using appropriate reference genes that are stably expressed under a variety of pathological and experimental conditions. No studies have validated specific reference genes in canine osteosarcoma (OS). Previous gene expression studies involving canine OS have used one or two reference genes to normalize gene expression. This study aimed to validate a panel of reference genes commonly used for normalization of canine OS gene expression data using the geNorm algorithm. qPCR analysis of nine canine reference genes was performed on 40 snap-frozen primary OS tumors and seven cell lines.

    RESULTS: Tumors with a variety of clinical and pathological characteristics were selected. Gene expression stability and the optimal number of reference genes for gene expression normalization were calculated. RPS5 and HNRNPH were highly stable among OS cell lines, while RPS5 and RPS19 were the best combination for primary tumors. Pairwise variation analysis recommended four and two reference genes for optimal normalization of the expression data of canine OS tumors and cell lines, respectively.

    CONCLUSIONS: Appropriate combinations of reference genes are recommended to normalize mRNA levels in canine OS tumors and cell lines to facilitate standardized and reliable quantification of target gene expression, which is essential for investigating key genes involved in canine OS metastasis and for comparative biomarker discovery.

    Matched MeSH terms: Real-Time Polymerase Chain Reaction/methods; Real-Time Polymerase Chain Reaction/veterinary
  3. Nikzad S, Tan SG, Yong Seok Yien C, Ng J, Alitheen NB, Khan R, et al.
    J Med Primatol, 2014 Dec;43(6):433-44.
    PMID: 24930735 DOI: 10.1111/jmp.12130
    The genetic diversity and structure of long-tailed macaques (Macaca fascicularis) in Peninsular Malaysia, a widely used non-human primate species in biomedical research, have not been thoroughly characterized.
    Matched MeSH terms: Polymerase Chain Reaction
  4. Harun MS, Kuan CO, Selvarajah GT, Wei TS, Arshad SS, Hair Bejo M, et al.
    Virol J, 2013;10:329.
    PMID: 24209771 DOI: 10.1186/1743-422X-10-329
    BACKGROUND:
    Feline Infectious Peritonitis (FIP) is a lethal systemic disease, caused by the FIP Virus (FIPV); a virulent mutant of Feline Enteric Coronavirus (FECV). Currently, the viruses virulence determinants and host gene expressions during FIPV infection are not fully understood.

    METHODS:
    RNA sequencing of Crandell Rees Feline Kidney (CRFK) cells, infected with FIPV strain 79-1146 at 3 hours post infection (h.p.i), were sequenced using the Illumina next generation sequencing approach. Bioinformatic's analysis, based on Felis catus 2X annotated shotgun reference genome, using CLC bio Genome Workbench mapped both control and infected cell reads to 18899 genes out of 19046 annotated genes. Kal's Z test statistical analysis was used to analyse the differentially expressed genes from the infected CRFK cells. Real time RT-qPCR was developed for further transcriptional profiling of three genes (PD-1, PD-L1 and A3H) in infected CRFK cells and Peripheral Blood Mononuclear Cells (PBMCs) from healthy and FIP-diseased cats.

    RESULTS:
    Based on Kal's Z-test, with False Discovery Rate (FDR) <0.05 and >1.99 fold change on gene expressions, a total of 61 genes were differentially expressed by both samples, where 44 genes were up-regulated and the remainder were down-regulated. Most genes were closely clustered together, suggesting a homogeneous expression. The majority of the genes that were significantly regulated, were those associated with monocytes-macrophage and Th1 cell functions, and the regulation of apoptosis. Real time RT-qPCR developed focusing on 2 up-regulated genes (PD-L1 and A3H) together with an apoptosis associated gene PD-1 expressions in FIPV infected CRFK cells and in PBMCs from healthy and FIP diagnosed cats produced concordant results with transcriptome data.

    CONCLUSION:
    The possible roles of these genes, and their importance in feline coronaviruses infection, are discussed.
    Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/methods; Real-Time Polymerase Chain Reaction/methods
  5. Zokaeifar H, Balcázar JL, Saad CR, Kamarudin MS, Sijam K, Arshad A, et al.
    Fish Shellfish Immunol, 2012 Oct;33(4):683-9.
    PMID: 22659618 DOI: 10.1016/j.fsi.2012.05.027
    We studied the effect of two probiotic Bacillus subtilis strains on the growth performance, digestive enzyme activity, immune gene expression and disease resistance of juvenile white shrimp (Litopenaeus vannamei). A mixture of two probiotic strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU g(-1) feed to shrimp for eight weeks. In comparison to untreated control group, final weight, weight gain and digestive enzyme activity were significantly greater in shrimp fed BM5 and BM8 diets. Significant differences for specific growth rate (SGR) and survival were recorded in shrimp fed BM8 diet as compared with the control; however, no significant differences were recorded for food conversion ratio (FCR) among all the experimental groups. Eight weeks after the start of the feeding period, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 63.3%, whereas cumulative mortality of the shrimp that had been given probiotics was 20.0% with BM8 and 33.3% with BM5. Subsequently, real-time PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan-binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was significantly up-regulated (P 
    Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction/veterinary; Real-Time Polymerase Chain Reaction/veterinary
  6. Teh SH, Fong MY, Mohamed Z
    Genet Mol Biol, 2011 Jul;34(3):464-70.
    PMID: 21931521 DOI: 10.1590/S1415-47572011005000022
    The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo.
    Matched MeSH terms: Polymerase Chain Reaction
  7. Bhassu S, See LM, Hassan R, Siraj SS, Tan SG
    Mol Ecol Resour, 2008 Sep;8(5):983-5.
    PMID: 21585948 DOI: 10.1111/j.1755-0998.2008.02127.x
    Eight single locus microsatellite markers were developed to characterize the Malaysian giant freshwater prawn, Macrobrachium rosenbergii. These microsatellites were isolated from an enriched genomic library contained by using a 5'-anchored polymerase chain reaction technique. Primers were designed to flank the repeat sequences and subsequently used to characterize 30 unrelated individuals of the giant freshwater prawn. The polymerase chain reaction amplification products of these eight microsatellite loci were polymorphic with the number of alleles ranging from two to 10 alleles per locus while the levels of heterozygosity ranged from 0.6333 to 0.8667.
    Matched MeSH terms: Polymerase Chain Reaction
  8. Maruyama S, Minaguchi K, Takezaki N, Nambiar P
    Leg Med (Tokyo), 2008 May;10(3):160-2.
    PMID: 18180191 DOI: 10.1016/j.legalmed.2007.11.002
    Allele frequencies for 15 short tandem repeat (STR) loci D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA (AmpF/STR Identifiler PCR Amplification kit, PE Applied Biosystems) were obtained from a sample of 110 unrelated individuals from the Malay population living in and around Kuala Lumpur, Malaysia, and the characteristics of the population was compared with other East Asian populations.
    Matched MeSH terms: Polymerase Chain Reaction
  9. Ng YH, Subramaniam V, Lau YL
    Vet Parasitol, 2015 Nov 30;214(1-2):200-3.
    PMID: 26455572 DOI: 10.1016/j.vetpar.2015.09.032
    Sarcocystosis in meat-producing animals is a major cause of reduced productivity in many countries, especially those that rely on agriculture. Although several diagnostic methods are available to detect sarcocystosis, many are too time-consuming for routine use in abattoirs and meat inspection centers, where large numbers of samples need to be tested. This study aimed to compare the sensitivity of the methylene blue tissue preparation, unstained tissue preparation and nested PCR in the detection of sarcocysts in tissue samples. Approximately three-fold more sarcocysts were detected in methylene blue-stained tissue compared to unstained controls (McNemar's test: P<0.01). Test sensitivity was comparable to that of the gold standard for sarcocyst detection, nested polymerase chain reaction. These results suggest that methylene blue can be used in tissue compression as a rapid, safe, and inexpensive technique for the detection of ruminant sarcocystosis in abattoirs.
    Matched MeSH terms: Polymerase Chain Reaction
  10. Fulazzaky MA, Abdullah S, Salim MR
    Data Brief, 2016 Jun;7:834-8.
    PMID: 27077083 DOI: 10.1016/j.dib.2016.03.058
    The goal of this study was to identify the biosurfactant-producing bacteria isolated from agro-food industrial effluet. The identification of the potential bacterial strain using a polymerase chain reaction of the 16S rRNA gene analysis was closely related to Serratia marcescens with its recorded strain of SA30 "Fundamentals of mass transfer and kinetics for biosorption of oil and grease from agro-food industrial effluent by Serratia marcescens SA30" (Fulazzaky et al., 2015) [1]; however, many biochemical tests have not been published yet. The biochemical tests of biosurfactant production, haemolytic assay and cell surface hydrophobicity were performed to investigate the beneficial strain of biosurfactant-producing bacteria. Here we do share data collected from the biochemical tests to get a better understanding of the use of Serratia marcescens SA30 to degrade oil, which contributes the technical features of strengthening the biological treatment of oil-contaminated wastewater in tropical environments.
    Matched MeSH terms: Polymerase Chain Reaction
  11. Hanapi UK, Desa MN, Ismail A, Mustafa S
    J Food Sci Technol, 2015 Jul;52(7):4166-75.
    PMID: 26139881 DOI: 10.1007/s13197-014-1459-7
    A Common Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of four groups of animal (pig, ruminant, avian and rabbit). This method demonstrated higher sensitivity and efficiency than the conventional multiplex PCR. In this approach, a common forward primer was designed in the 5' end of a homologous region of mitochondrial NADH dehyrogenase subunit 4 (Nad 4) gene sequences of all the animal groups. Specific adapter reverse primers were designed by adding an adapter sequence at the 5' end. The same adapter sequence was used as the common adapter reverse primer. The primers generated specific fragments of 267, 370, 504, and 548 bp lengths for pig, ruminant, avian and rabbit meats, respectively. The use of adapter sequence at the 5' end of the common adapter reverse primers increased the efficiency of the amplification and the application of a common forward primer solved the complexity in multiplex PCR system. Bands of specific amplification can be detected in the PCR assays containing as low as 10(-6) μM of adapter reverse primer. This result indicated that the sensitivity was tremendously increased as compared to the conventional multiplex PCR (10(-3) μM). CP-M-PCR detection limit of the DNA samples was 0.1 ng for the four groups of meats. CP-M-PCR has greatly improved the sensitivity and efficiency of the PCR system for a more reliable and accurate outcome than conventional multiplex PCR system.
    Matched MeSH terms: Multiplex Polymerase Chain Reaction
  12. Tan GK, Tee SF, Tang PY
    Genet Mol Biol, 2015 May;38(2):138-46.
    PMID: 26273215 DOI: 10.1590/S1415-4757382220140142
    Dystrobrevin binding protein 1 (DTNBP1) gene is pivotal in regulating the glutamatergic system. Genetic variants of the DTNBP1 affect cognition and thus may be particularly relevant to schizophrenia. We therefore evaluated the association of six single nucleotide polymorphisms (SNPs) with schizophrenia in a Malaysian population (171 cases; 171 controls). Associations between these six SNPs and schizophrenia were tested in two stages. Association signals with p < 0.05 and minor allele frequency > 0.05 in stage 1 were followed by genotyping the SNPs in a replication phase (stage 2). Genotyping was performed with sequenced specific primer (PCR-SSP) and restriction fragment length polymorphism (PCR-RFLP). In our sample, we found significant associations between rs2619522 (allele p = 0.002, OR = 1.902, 95%CI = 1.266 - 2.859; genotype p = 0.002) and rs2619528 (allele p = 0.008, OR = 1.606, 95%CI = 1.130 - 2.281; genotype p = 6.18 × 10(-5)) and schizophrenia. Given that these two SNPs may be associated with the pathophysiology of schizophrenia, further studies on the other DTNBP1 variants are warranted.
    Matched MeSH terms: Polymerase Chain Reaction
  13. Khoo AS, Balraj P, Rachedi A, Chin CN, Volpi L
    Hum Mutat, 1999 Nov;14(5):448.
    PMID: 10533073
    Matched MeSH terms: Polymerase Chain Reaction
  14. Norazah A, Zainuldin MT, Kamel AGM, Kamaliah MN, Taha AM
    Med J Malaysia, 2001 Mar;56(1):4-9.
    PMID: 11503295
    The detection of Vibrio cholerae 01 from the aquatic environment of Daro and Bintulu in Sarawak was carried out following an outbreak of cholera. Conventional culture methods and detection of ctx gene by polymerase chain reaction technique were carried out on 80 water samples. Only one sample was positive by culture methods while 8 were positive by PCR. DNA finger printing by pulsed-field gel electrophoresis showed that the clinical isolates in Daro and Bintulu were genetically identical while the environmental isolate was closely related. Recovery of Vibrio cholerae by culture method is poor and newer methods of detection should be developed.
    Matched MeSH terms: Polymerase Chain Reaction
  15. Lee J, Hughes T, Lee MH, Field H, Rovie-Ryan JJ, Sitam FT, et al.
    Ecohealth, 2020 09;17(3):406-418.
    PMID: 33226526 DOI: 10.1007/s10393-020-01503-x
    The legal and illegal trade in wildlife for food, medicine and other products is a globally significant threat to biodiversity that is also responsible for the emergence of pathogens that threaten human and livestock health and our global economy. Trade in wildlife likely played a role in the origin of COVID-19, and viruses closely related to SARS-CoV-2 have been identified in bats and pangolins, both traded widely. To investigate the possible role of pangolins as a source of potential zoonoses, we collected throat and rectal swabs from 334 Sunda pangolins (Manis javanica) confiscated in Peninsular Malaysia and Sabah between August 2009 and March 2019. Total nucleic acid was extracted for viral molecular screening using conventional PCR protocols used to routinely identify known and novel viruses in extensive prior sampling (> 50,000 mammals). No sample yielded a positive PCR result for any of the targeted viral families-Coronaviridae, Filoviridae, Flaviviridae, Orthomyxoviridae and Paramyxoviridae. In the light of recent reports of coronaviruses including a SARS-CoV-2-related virus in Sunda pangolins in China, the lack of any coronavirus detection in our 'upstream' market chain samples suggests that these detections in 'downstream' animals more plausibly reflect exposure to infected humans, wildlife or other animals within the wildlife trade network. While confirmatory serologic studies are needed, it is likely that Sunda pangolins are incidental hosts of coronaviruses. Our findings further support the importance of ending the trade in wildlife globally.
    Matched MeSH terms: Polymerase Chain Reaction
  16. Nur Syahira Mohammad, Zaidah Zainal Ariffin
    Science Letters, 2020;14(2):15-23.
    MyJurnal
    Fungi is known to produce a wide range of biologically active metabolites and enzymes. Enzymes produced by fungi are utilized in food and pharmaceutical industries because of their rich enzymatic profile. Filamentous fungi are particularly interesting due to their high production of extracellular enzymes which has a large industrial potential. The aim of this study is to isolate potential soil fungi species that are able to produce functional enzymes for industries. Five Aspergillus species were successfully isolated from antibiotic overexposed soil (GPS coordinate of N3.093219 E101.40269) by standard microbiological method. The isolated fungi were identified via morphological observations and molecular tools; polymerase chain reactions, ITS 1 (5’- TCC GTA GGT GAA CCT GCG G3’) forward primer and ITS 4 (5’-TCC TCC GCT TAT TGA TAT GC-3’) reverse primer. The isolated fungi were identified as Aspergillus sydowii strain SCAU066, Aspergillus tamarii isolate TN-7, Aspergillus candidus strain KUFA 0062, Aspergillus versicolor isolate BAB-6580, and Aspergillus protuberus strain KAS 6024. Supernatant obtained via submerged fermentation of the isolated fungi in potato dextrose broth (PDB) and extracted via centrifugation was loaded onto specific media to screen for the production of xylanolytic, cellulolytic and amylolytic enzymes. The present findings indicate that Aspergillus sydowii strain SCAU066 and Aspergillus versicolor isolate BAB-6580 have great potential as an alternative source of xylanolytic, cellulolytic and amylolytic enzymes.
    Matched MeSH terms: Polymerase Chain Reaction
  17. Rupert R, Lie GJCW, John DV, Annammala KV, Jani J, Rodrigues KF
    Data Brief, 2020 Dec;33:106351.
    PMID: 33072827 DOI: 10.1016/j.dib.2020.106351
    The data provided in the article includes the sequence of bacterial 16S rRNA gene from a high conservation value forest, logged forest, rubber plantation and oil palm plantation collected at Kelantan river basin. The logged forest area was previously notified as a flooding region. The total gDNA of bacterial community was amplified via polymerase chain reaction at V3-V4 regions using a pair of specific universal primer. Amplicons were sequenced on Illumina HiSeq paired-end platform to generate 250 bp paired-end raw reads. Several bioinformatics tools such as FLASH, QIIME and UPARSE were used to process the reads generated for OTU analysis. Meanwhile, R&D software was used to construct the taxonomy tree for all samples. Raw data files are available at the Sequence Read Archive (SRA), NCBI and data information can be found at the BioProject and BioSample, NCBI. The data shows the comparison of bacterial community between the natural forest and different land uses.
    Matched MeSH terms: Polymerase Chain Reaction
  18. Jainlabdin MH, Batra A, Sánchez Paredes E, Hernández Hernández F, Fu G, Tovar-Torres J
    Sci Rep, 2019 10 11;9(1):14692.
    PMID: 31604994 DOI: 10.1038/s41598-019-51198-6
    Invasive candidiasis is one of the most common nosocomial fungal infections worldwide. Delayed implementation of effective antifungal treatment caused by inefficient Candida diagnosis contributes to its notoriously high mortality rates. The availability of better Candida diagnostic tools would positively impact patient outcomes. Here, we report on the development of a single-tube, dual channel pentaplex molecular diagnostic assay based on Multiplex Probe Amplification (MPA) technology. It allows simultaneous identification of C. auris, C. glabrata and C. krusei, at species-level as well as of six additional albicans and non-albicans pathogenic Candida at genus level. The assay overcomes the one-channel one-biomarker limitation of qPCR-based assays. Assay specificities are conferred by unique biomarker probe pairs with characteristic melting temperatures; post-amplification melting curve analysis allows simple identification of the infectious agent. Alerting for the presence of C. auris, the well-characterised multi-drug resistant outbreak strain, will facilitate informed therapy decisions and aid antifungal stewardship. The MPA-Candida assay can also be coupled to a pan-Fungal assay when differentiation between fungal and bacterial infections might be desirable. Its multiplexing capacity, detection range, specificity and sensitivity suggest the potential use of this novel MPA-Candida assay in clinical diagnosis and in the control and management of hospital outbreaks.
    Matched MeSH terms: Multiplex Polymerase Chain Reaction/methods*; Real-Time Polymerase Chain Reaction/methods
  19. Tobe SS, Swaran YC, Dennany L, Sibbing U, Schulze Johann K, Welch L, et al.
    Int J Legal Med, 2017 Jan;131(1):87-94.
    PMID: 27832353 DOI: 10.1007/s00414-016-1461-x
    Sexual assault samples are some of the most common samples encountered in forensic analysis. These samples can require a significant time investment due to differential extraction processes. We report on the first record of successful direct amplification of semen for STR analysis. Neat seminal fluid, dilutions ranging from 1:5 to 1:160 and GEDNAP samples were successfully amplified using a direct method. A mild differential isolation technique to enrich spermatozoa was developed and successfully implemented to separate and directly amplify a mixture of semen and female epithelial cells. Aliquots of samples subjected to the differential isolation protocol were stained with Haemotoxylin and Eosin for sperm scoring. Samples stained after PCR showed a complete lack of intact spermatozoa demonstrating that the cells are lysed during the PCR process. This paper demonstrates the potential to incorporate direct PCR in cases of sexual assault to more rapidly obtain results and achieve a higher sensitivity.
    Matched MeSH terms: Polymerase Chain Reaction
  20. Radzi AM, Hun KS, Kong N, Yahaya N
    Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease exhibiting extensive clinical heterogeneity. Genetic factors and immune dysregulation play important roles in its development. Apoptosis is a physiologic process that regulates normal homeostasis. It is likely to contribute to the pathogenesis of autoimmune diseases by impairing elimination of autoreactive T and B cells. Apo-1/Fas which is a transmembrane protein mediates apoptosis and is a member of the tumour necrosis factor/nerve growth factor receptor family. It transduces the apoptotic signal into susceptible target cells. Recent studies have focused on the apoptosis mediated by these proteins in the causation of several autoimmune disorders including SLE. Aim: To determine the frequency of Apo-1/Fas promoter gene polymorphism in patients with systemic lupus erythematosus and healthy controls and to investigate its role in the susceptibility of SLE in a cohort of Malaysian Chinese SLE patients Materials and methods: 107 Chinese patients and 60 matched controls were genotyped using polymerase chain reaction (PCR) amplification followed by MvaI restriction enzyme digestion. The MvaI RFLP is located at the -670 position from the transcription starting site and results from an A→G substitution which alters the MvaI restriction site. Results: G/G genotype was found in 25% of patients and controls while the A/A genotype in 23% and 31.6% of patients and controls respectively. Heterozygous form was noted in 43% of the normal population compared to 51% in SLE patients. There was also no significant difference in the allele frequencies of G and A in both groups studied. Conclusion: We suggest that polymorphism of the Apo-1/Fas promoter gene does not play a role in disease susceptibility in Chinese patients with SLE. © 2008 Japan International Cultural Exchange Foundation.
    Matched MeSH terms: Polymerase Chain Reaction
Filters
Contact Us

Please provide feedback to Administrator (afdal@afpm.org.my)

External Links