Search

[Objective]Fluorescent capillary electrophoresis, for its high detection throughput and resolution, was in extensive applications in various scenarios, including individual identification, variety discrimination, and species identification. The tailed-sequence offers an efficient solution for the extensive application of the fluorescent capillary electrophoresis platforms. To address the limitation of existing tailed-sequences in meeting the demands of multi-species applications, this study developed an efficient Universal Tailed-Sequence (UTS)design tool based on encoding translation technology. Utilizing this tool, UTS were designed to construct a versatile PCR system and protocol suitable for multiple crops, thereby enhancing the throughput and flexibility of fluorescent electrophoresis.[Method]An efficient UTS design tool was designed based on encoding translation technology and 3 755 commonly used Chinese characters were encoding translated with filtering criteria including GC content, hairpin, and the self-dimer annealing temperature. The BLAST tool was used to assess the homology of UTS generated by the design tool on various biological genomes. The UTS was experimentally evaluated on crops such as maize, tomatoes, peppers, and watermelons, constructing a species-universal experimental system and protocol.[Result]The design tool, through encoding translation and screening, generated a total of 7 436 833 high-quality candidate UTS, accounting for 52.74% of all character combinations. Six selected UTS were demonstrated better specificity across 20 crop genomes by BLAST results compared to M13. With optimization of the general primer amplification procedure, the success rate of general primer amplification across multiple species reached or exceeded 95%, showing strong stability.[Conclusion]This study utilized encoding translation technology to develop UTS and paired them with a species-universal amplification system and protocol, which may provide a high-throughput, cost-effective universal detection method for fluorescent electrophoresis. [ABSTRACT FROM AUTHOR]

Coconut (Cocos nucifera) sap is one of the natural drinks, being traditionally tapped from unopened inflorescences of the coconut palms. In the present study, microbial quality of coconut sap that was collected using three methods; Treatment 1: application of hal bark (Vateria copallifera) to the 4L clay pots (TM), Treatment 2: Novel sap collection method (NSM), Treatment 3: application of 5g of hal bark in to 30 cm X 50 cm polythene bag in 4L clay pots (MTM) were evaluated. Sap was collected for 12 h duration and Total Plate Count (TPC) and Yeast and Mold Count (YMC) were determined. Colonies were isolated and preserved. DNA extractions of microbes were done by CTAB (Cetyl trimethylammonium bromide) method with modification. ITS1 forward and ITS4 reverse primers were used to identify yeast species and 27 forward and 1492 reverse primers were used for the bacterial species in Polymerase Chain Reaction (PCR). Amplified products were separated using 1.5 % agarose gel and purified DNA was sent into Macrogene Korea for sequencing. Four types (A, B, C and D) of distinct microbial colonies were isolated from the differently collected coconut sap samples. DNA homology data revealed that, A is Naumannella halotolerans only presented in NSM. B and C Serratia marcescens, are Achromobacter xylosoxidans contained in TM and MTM. Saccharomyces cerevisiae (D) was found in all the collection system. The collection method affects for the microbial quality and quantity of unfermented coconut sap. [ABSTRACT FROM AUTHOR]

Copyright of Serbian Dental Journal / Stomatološki Glasnik Srbije is the property of National Library of Serbia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Jurnal Fitopatologi Indonesia is the property of IPB University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Environmental DNA (eDNA) metabarcoding is a powerful tool for monitoring biodiversity in natural ecosystems. The accuracy of eDNA metabarcoding relies heavily on the PCR primers that amplify target sequences. For vertebrates, most available primers are designed for particular groups. There have been inadequate attempts to design “universal” primers applicable to all vertebrates. Here, we developed three new universal primer sets (V12S-U, V16S-U, and VCOI-U) targeting mitochondrial 12S, 16S, and COI genes, respectively. They work for all vertebrate groups, with amplification length of 200-250 bp, facilitating the use of short-read sequencing platforms. We evaluated and compared the species detection ability of our primer sets and seven previously published ones through both in-silico PCR and experiment tests with mock DNA and zoo eDNA. The species detection success of the new primer sets is over 90%, showing better performance than previously published primer sets. We also tested our new primers with water eDNA samples of natural environments, and detected a total of 895 vertebrate OTUs (BLAST identity ≥ 0.95), comprising 182 species, 195 genera, and 94 families. Our results indicated that utilizing multiple markers can achieve better species detection than using only a single marker, which is especially important for monitoring large biological communities like vertebrates. The new primer sets enable researchers to detect the presence of species of different vertebrate groups in one eDNA metabarcoding survey, which simplifies the workflow and reduces the cost. It has the potential to serve as an alternative/complementary tool for future eDNA metabarcoding studies targeting vertebrates.

This study aims to compare the shear bond strength (SBS) of a direct resin composite to CAD-CAM resin composite blocks treated with different surface treatments: micromechanical, chemical or a combination of both. Eight CAD-CAM resin composite blocks, namely Brilliant Crios, Cerasmart 270, Vita Enamic, Grandio block, Katana Avencia, Lava Ultimate, Tetric CAD and Shofu Block HC were chosen. The micromechanical surface treatment protocols tested were hydrofluoric acid, polyacrylic acid or sandblasting, and the chemical one was a universal primer. These treated CAD-CAM blocks were tested to determine the SBS of a light-curing composite resin Z100 bonded to their surface. Two-way ANOVA followed by Tukey's post hoc test was used to investigate the difference in SBS. Failures were analyzed by Fisher's exact test. Bonding interfaces were examined by scanning electron microscopy. The micromechanical surface treatments give the highest SBS values: sandblasting appears to be the most efficient procedure for dispersed filler composite blocks, while hydrofluoric acid etching is preferable for polymer-infiltrated ceramic network (PICN) blocks. The use of universal primer does not improve SBS values on dispersed filler composite blocks. For PICN blocks, the use of universal primer significantly increases SBS values when combined with hydrofluoric acid etching. [ABSTRACT FROM AUTHOR]

Objective: The purpose of this study is to compare the bonding performance and mechanical properties of two different resin composite cements using simplified adhesive bonding strategies. Materials and methods: Shear bond strength of two resin composite cements (an adhesive cement: Panavia V5 [PV5] and a self‐adhesive cement: RelyX Universal [RUV]) to human enamel, dentin, and a variety of restorative materials (microfilled composite, composite, polymer‐infiltrated ceramic, feldspar ceramic, lithium disilicate and zirconia) was measured. Thermocycle aging was performed with selected material combinations. Results: For both cements, the highest shear bond strength to dentin was achieved when using a primer (PV5: 18.0 ± 4.2 MPa, RUV: 18.2 ± 3.3 MPa). Additional etching of dentin reduced bond strength for RUV (12.5 ± 4.9 MPa). On enamel, PV5 achieved the highest bond strength when the primer was used (18.0 ± 3.1 MPa), while for RUV etching of enamel and priming provided best results (21.2 ± 6.6 MPa). Shear bond strength of RUV to restorative materials was superior to PV5. Bonding to resin‐based materials was predominantly observed for RUV. Conclusions: While use of RUV with the selective‐etch technique is slightly more labor intensive than PV5, RUV (with its universal primer) displayed a high‐bonding potential to all tested restorative materials, especially to resin. Clinical significance: For a strong adhesion to the tooth substrate, PV5 (with its tooth primer) is to be preferred because etching with phosphoric acid is not required. However, when using a wide range of varying restorative materials, RUV with its universal primer seems to be an adequate option. [ABSTRACT FROM AUTHOR]

Abstract Background Flaviviruses are representative arboviruses carried by arthropods and/or vertebrates; these viruses can pose a public health concern in many countries. By contrast, it is known that a novel virus group called insect-specific flaviviruses (ISFs) also infects arthropods, although no such virus has yet been isolated from vertebrates. The characteristics of ISFs, which affect replication of human-pathogenic flaviviruses within co-infected mosquito cells or mosquitoes without affecting the mosquitoes themselves, mean that we should pay attention to both ISFs and human-pathogenic flaviviruses, despite the fact that ISFs appear not to be directly hazardous to human health. To assess the risk of diseases caused by flaviviruses, and to better understand their ecology, it is necessary to know the extent to which flaviviruses are harbored by arthropods. Methods We developed a novel universal primer for use in a PCR-based system to detect a broad range of flaviviruses. We then evaluated its performance. The utility of the novel primer pair was evaluated in a PCR assay using artificially synthesized oligonucleotides derived from a template viral genome sequence. The utility of the primer pair was also examined by reverse transcription PCR (RT-PCR) using cDNA templates prepared from virus-infected cells or crude supernatants prepared from virus-containing mosquito homogenates. Results The novel primer pair amplified the flavivirus NS5 sequence (artificially synthesized) in all samples tested (six species of flavivirus that can cause infectious diseases in humans, and flaviviruses harbored by insects). In addition, the novel primer pair detected viral genomes in cDNA templates prepared from mosquito cells infected with live flavivirus under different infectious conditions. Finally, the viral genome was detected with high sensitivity in crude supernatants prepared from pooled mosquito homogenates. Conclusion This PCR system based on a novel primer pair makes it possible to detect arthropod-borne flaviviruses worldwide (the primer pair even detected viruses belonging to different genetic subgroups). As such, an assay based on this primer pair may help to improve public health and safety, as well as increase our understanding of flavivirus ecology.

Purpose: To evaluate the effects of tribochemical silica coating and different 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-containing primers on the shear bond strength (SBS) of orthodontic metal brackets to yttriumstabilized tetragonal zirconia polycrystal (Y-TZP) surfaces. Materials and Methods: One hundred ninety polished Y-TZP specimens were randomly assigned to 19 groups (n = 10): 30 specimens were used for surface analyses after polishing with 600-grit silicon carbide paper, airborneparticle abrasion with 50-µm alumina (A), or tribochemical silica coating (CoJet [C]); 160 specimens were used in SBS testing of orthodontic metal brackets to Y-TZP after alumina airborne-particle abrasion or tribochemical silica coating and application of either ESPE-Sil (S) (ASn, ASa, CSn, CSa), Alloy Primer (AP) (AAPn, AAPa, CAPn, CAPa), Clearfil Ceramic Primer (CP) (ACPn, ACPa, CCPn, CCPa), or Scotchbond Universal (U) (AUn, AUa, CUn, CUa) and either stored in water for 24 h (non-aged, n) or thermocycled 5000 times (aged, a). The surface analyses and SBSs were statistically analyzed with ANOVA and Tukey's tests. Results: Both mechanically treated surfaces had significantly greater surface roughness and surface free energy than did the polished surfaces. The type of primer and aging significantly affected the bond strength. Among the thermocycled specimens, the AAPa, AUa, and CCPa groups showed the greatest SBS. Conclusion: After alumina airborne-particle abrasion, the application of Alloy Primer, Clearfil Ceramic Primer, or Scotchbond Universal provided stable bonding to Y-TZP ceramics. After tribochemical silica coating, however, only Clearfil Ceramic Primer produced a durable bond to Y-TZP ceramics. [ABSTRACT FROM AUTHOR]

Cucurbit crops belonging to the family Cucurbitaceae are one the most important horticultural crops worldwide and cultivated widely in Iran. To aim detection and phylogenetic analysis of potyvirus members, during 2015-2016, 215 leaf samples from cucumber, melon, watermelon, and squash symptomatic with deforming and reduction in leaf size, blistering, mild and severe mosaic, fruit deformation, and stunting were collected from 10 major cucurbits-growing locations in West Azerbaijan province. Following mechanical inoculation of Cucurbita pepo as an indicator plant with symptomatic cucurbit crop extracts, a variety of symptoms developed on the plants. Then, total RNA extracted from 70 symptomatic leaf samples, and partial RNA-dependent RNA polymerase (NIb) (350 bp) was amplified using universal primer pairs (NIb2F/NIb3R) by reverse transcription-polymerase chain reaction (RT-PCR). The infection by potyviruses in 38.75% of the samples was confirmed based on the result. The five PCR products belonging to different hosts at the expected size (350bp) were sequenced. One sample (Ir-Na: MH491979) was determined to be infected by the Watermelon mosaic virus (WMV) whereas four samples (Ir-Ma:MH491979, Ir-Ng:MH491980, Ir-Pi:MH491981, and Ir-Ur:MH491982) infected by Zucchini yellow mosaic virus (ZYMV). In the phylogenetic analysis based on Maximum-likelihood, three ZYMV isolates (Ir-Ng, Ir-Pi, and Ir-Ur) were placed in subgroup I whereas one isolate (Ir-Ma) was placed in subgroup II belonging to group A. Also, WMV isolate (Ir- Na) was placed in CL phylogroup.

Background: Recent studies have reported that melanoma antigen (MAGE) gene is expressed in a variety of cancers and testicular tissues. The expression of MAGE-A genes could be used for biomarkers with high tumor specificity; however, there is still a lack of data on most solid tumors. The objective of this study was to construct novel universal primers for detecting the mRNA of MAGE A1-10 genes in lung cancer patients. Method: We conducted this cross-sectional study in 2017 at Dr. Soetomo General Academic Hospital, Surabaya, Indonesia. The specimens were a testicular tissue and 15 core biopsies of lung cancer tissues. We designed the universal primers to bind the mRNA of MAGE A1, A2, A2B, A3, A4, A5, A6, A8, A9, A9B, and A10 regions; the assay was performed by nested PCR and continued by direct sequencing. Results: Using the universal primer MAGE A1-10, the PCR was able to detect the MAGE A mRNA of 10 subtypes of MAGE A from testicular and lung cancer tissues. The sequences analysis of individual MAGE A1-10 showed the same homology with MAGE A from GenBank data. Among the 15 lung cancer patients, 13/15 (86.67%) tested positive for GAPDH; subsequently, they were considered for MAGE-A gene detection; while, those testing negative for GAPDH were excluded. The PCR results showed that 12/13 (92.31%) had positive MAGE A1-10 tests and 3/13 (23.08%) tested positive for MAGE A1-6. Conclusion: This finding showed that the novel universal primers could be applied as a new tool for detecting MAGE A1-10 expression in cancer cells.

Yellow leaf curl disease in chili pepper has been reported in Bali Island since the early 2012. Research was conducted to identify the virus causing this disease and disease distribution in Bali. Field survey was carried out to observe disease intensity and to collect field samples from several chili pepper growing areas in Bali (Karangasem, Bangli, Tabanan, and Gianyar). Begomovirus identification from field samples was then conducted by polymerase chain reaction method using universal primers SPG1/SPG2, followed by an analysis of the amplified target DNA sequences. The incidence of pepper yellow leaf curl disease reached 100% at all sites and disease severity reached 18%−87%. Begomovirus specific DNA fragment measuring 912 bp was successfully amplified from 12 field samples. Sequence analysis of DNA fragments showed the highest homology with Pepper yellow leaf curl Indonesia virus (PYLCIV). Further phylogenetic analysis confirmed the relationship between PYLCIV isolates from Bali and various PYLCIV isolates from Indonesia.

Abstract Background Amplification of small subunit (SSU) rRNA genes with universal primers is a common method used to assess microbial populations in various environmental samples. However, owing to limitations in coverage of these universal primers, some microorganisms remain unidentified. The present study aimed to establish a method for amplifying nearly full-length SSU rRNA gene sequences of previously unidentified prokaryotes, using newly designed targeted primers via primer evaluation in meta-transcriptomic datasets. Methods Primer binding regions of universal primer 8F/Arch21F for bacteria or archaea were used for primer evaluation of SSU rRNA sequences in meta-transcriptomic datasets. Furthermore, targeted forward primers were designed based on SSU rRNA reads from unclassified groups unmatched with the universal primer 8F/Arch21F, and these primers were used to amplify nearly full-length special SSU rRNA gene sequences along with universal reverse primer 1492R. Similarity and phylogenetic analysis were used to confirm their novel status. Results Using this method, we identified unclassified SSU rRNA sequences that were not matched with universal primer 8F and Arch21F. A new group within the Asgard superphylum was amplified by the newly designed specific primer based on these unclassified SSU rRNA sequences by using mudflat samples. Conclusion We showed that using specific primers designed based on universal primer evaluation from meta-transcriptomic datasets, identification of novel taxonomic groups from a specific environment is possible.

Background: Flaviviruses are representative arboviruses carried by arthropods and/or vertebrates; these viruses can pose a public health concern in many countries. By contrast, it is known that a novel virus group called insect-specific flaviviruses (ISFs) also infects arthropods, although no such virus has yet been isolated from vertebrates. The characteristics of ISFs, which affect replication of human-pathogenic flaviviruses within co-infected mosquito cells or mosquitoes without affecting the mosquitoes themselves, mean that we should pay attention to both ISFs and human-pathogenic flaviviruses, despite the fact that ISFs appear not to be directly hazardous to human health. To assess the risk of diseases caused by flaviviruses, and to better understand their ecology, it is necessary to know the extent to which flaviviruses are harbored by arthropods. Methods: We developed a novel universal primer for use in a PCR-based system to detect a broad range of flaviviruses. We then evaluated its performance. The utility of the novel primer pair was evaluated in a PCR assay using artificially synthesized oligonucleotides derived from a template viral genome sequence. The utility of the primer pair was also examined by reverse transcription PCR (RT-PCR) using cDNA templates prepared from virus-infected cells or crude supernatants prepared from virus-containing mosquito homogenates. Results: The novel primer pair amplified the flavivirus NS5 sequence (artificially synthesized) in all samples tested (six species of flavivirus that can cause infectious diseases in humans, and flaviviruses harbored by insects). In addition, the novel primer pair detected viral genomes in cDNA templates prepared from mosquito cells infected with live flavivirus under different infectious conditions. Finally, the viral genome was detected with high sensitivity in crude supernatants prepared from pooled mosquito homogenates. Conclusion: This PCR system based on a novel primer pair makes it possible to detect arthropod-borne flaviviruses worldwide (the primer pair even detected viruses belonging to different genetic subgroups). As such, an assay based on this primer pair may help to improve public health and safety, as well as increase our understanding of flavivirus ecology. [ABSTRACT FROM AUTHOR]

This study aims to compare the shear bond strength (SBS) of a direct resin composite to CAD-CAM resin composite blocks treated with different surface treatments: micromechanical, chemical or a combination of both. Eight CAD-CAM resin composite blocks, namely Brilliant Crios, Cerasmart 270, Vita Enamic, Grandio block, Katana Avencia, Lava Ultimate, Tetric CAD and Shofu Block HC were chosen. The micromechanical surface treatment protocols tested were hydrofluoric acid, polyacrylic acid or sandblasting, and the chemical one was a universal primer. These treated CAD-CAM blocks were tested to determine the SBS of a light-curing composite resin Z100 bonded to their surface. Two-way ANOVA followed by Tukey’s post hoc test was used to investigate the difference in SBS. Failures were analyzed by Fisher’s exact test. Bonding interfaces were examined by scanning electron microscopy. The micromechanical surface treatments give the highest SBS values: sandblasting appears to be the most efficient procedure for dispersed filler composite blocks, while hydrofluoric acid etching is preferable for polymer-infiltrated ceramic network (PICN) blocks. The use of universal primer does not improve SBS values on dispersed filler composite blocks. For PICN blocks, the use of universal primer significantly increases SBS values when combined with hydrofluoric acid etching.

Recent coronavirus (CoV) outbreaks, including that of Middle East respiratory syndrome (MERS), have presented a threat to public health worldwide. A primary concern in these outbreaks is the extent of mutations in the CoV, and the content of viral variation that can be determined only by whole genome sequencing (WGS). We aimed to develop a time efficient WGS protocol, using universal primers spanning the entire MERS-CoV genome. MERS and synthetic Neoromicia capensis bat CoV genomes were successfully amplified using our developed PCR primer set and sequenced with MinION. All experimental and analytical processes took 6 h to complete and were also applied to synthetic animal serum samples, wherein the MERS-CoV genome sequence was completely recovered. Results showed that the complete genome of MERS-CoV and related variants could be directly obtained from clinical samples within half a day. Consequently, this method will contribute to rapid MERS diagnosis, particularly in future CoV epidemics.

Cephalopods are pivotal components of marine food webs, but biodiversity studies are hampered by challenges to sample these agile marine molluscs. Metabarcoding of environmental DNA (eDNA) is a potentially powerful technique to study oceanic cephalopod biodiversity and distribution but has not been applied thus far. We present a novel universal primer pair for metabarcoding cephalopods from eDNA, Ceph18S (Forward: 5′-CGC GGC GCT ACA TAT TAG AC-3′, Reverse: 5′-GCA CTT AAC CGA CCG TCG AC-3′). The primer pair targets the hypervariable region V2 of the nuclear 18S rRNA gene and amplifies a relatively short target sequence of approximately 200 bp in order to allow the amplification of degraded DNA. In silico tests on a reference database and empirical tests on DNA extracts from cephalopod tissue estimate that 44–66% of cephalopod species, corresponding to about 310–460 species, can be amplified and identified with this primer pair. A multi-marker approach with the novel Ceph18S and two previously published cephalopod mitochondrial 16S rRNA primer sets targeting the same region (Jarman et al. 2006 Mol. Ecol. Notes. 6, 268–271; Peters et al. 2015 Mar. Ecol. 36, 1428–1439) is estimated to amplify and identify 89% of all cephalopod species, of which an estimated 19% can only be identified by Ceph18S. All sequences obtained with Ceph18S were submitted to GenBank, resulting in new 18S rRNA sequences for 13 cephalopod taxa.

Deer species has both scientific research and economic value, and half of these species, however, are listed as endangered animals. For the conservation purpose, we designed a novel universal deer-specific PCR primer pair based on an evolutionarily conservative coding sequence (i.e., CEP295NL gene) across some deer species. This primer pair was successfully amplified and sequenced, showing around ~ 540 bp in cervids. Validation results showed that it can be utilized to develop a reliable and simple diagnostic tool for distinguishing other closely related species, as well as possibly interspecific identification amongst cervids. [ABSTRACT FROM AUTHOR]

Recent coronavirus (CoV) outbreaks, including that of Middle East respiratory syndrome (MERS), have presented a threat to public health worldwide. A primary concern in these outbreaks is the extent of mutations in the CoV, and the content of viral variation that can be determined only by whole genome sequencing (WGS). We aimed to develop a time efficient WGS protocol, using universal primers spanning the entire MERS-CoV genome. MERS and synthetic Neoromicia capensis bat CoV genomes were successfully amplified using our developed PCR primer set and sequenced with MinION. All experimental and analytical processes took 6 h to complete and were also applied to synthetic animal serum samples, wherein the MERS-CoV genome sequence was completely recovered. Results showed that the complete genome of MERS-CoV and related variants could be directly obtained from clinical samples within half a day. Consequently, this method will contribute to rapid MERS diagnosis, particularly in future CoV epidemics. [ABSTRACT FROM AUTHOR]

Background: Recent studies have reported that melanoma antigen (MAGE) gene is expressed in a variety of cancers and testicular tissues. The expression of MAGEA genes could be used for biomarkers with high tumor specificity; however, there is still a lack of data on most solid tumors. The objective of this study was to construct novel universal primers for detecting the mRNA of MAGE A1-10 genes in lung cancer patients. Method: We conducted this cross-sectional study in 2017 at Dr. Soetomo General Academic Hospital, Surabaya, Indonesia. The specimens were a testicular tissue and 15 core biopsies of lung cancer tissues. We designed the universal primers to bind the mRNA of MAGE A1, A2, A2B, A3, A4, A5, A6, A8, A9, A9B, and A10 regions; the assay was performed by nested PCR and continued by direct sequencing. Results: Using the universal primer MAGE A1-10, the PCR was able to detect the MAGE A mRNA of 10 subtypes of MAGE A from testicular and lung cancer tissues. The sequences analysis of individual MAGE A1-10 showed the same homology with MAGE A from GenBank data. Among the 15 lung cancer patients, 13/15 (86.67%) tested positive for GAPDH; subsequently, they were considered for MAGEA gene detection; while, those testing negative for GAPDH were excluded. The PCR results showed that 12/13 (92.31%) had positive MAGE A1-10 tests and 3/13 (23.08%) tested positive for MAGE A1-6. Conclusion: This finding showed that the novel universal primers could be applied as a new tool for detecting MAGE A1-10 expression in cancer cells. [ABSTRACT FROM AUTHOR]

One of the most important diseases of pea is caused by Pea seed-borne mosaic virus (PSbMV), which has a relatively wide host range. Since there are few varieties with resistance against the virus, and spraying insecticides is not very effective, the determination of the disease and the pathogen in the seeds is very important. Inoculum prepared from pea seeds showing typical virus symptoms caused very mild symptoms on Chenopodium amaranticolor and C. quinoa, but several chlorotic/necrotic lesions on bean (Phaseolus vulgaris) cv. Scarlet, and systemic symptoms with mosaic and curling of top leaves on bean cv. Maxidor. The detection of the virus was carried out by PCR using universal primers and virus sequence analysis. According to the phylogenetic analysis the PSbMV isolate identified in Hungary belongs to the pathotype P1 and associated with the cluster 2 isolates. [ABSTRACT FROM AUTHOR]

BACKGROUND: Rapid and accurate species identification is not only important for biodiversity studies and pest quarantine and management, but in some cases may also influence the results of international trade negotiations. In this study, we developed a rapid species identification system for insects. RESULTS: A universal DNA mini‐barcode primer pair was designed to target ∼ 120 bp of the mitochondrial 16S rDNA gene. This primer set can amplify the targeted region from all 300 species of 26 insect orders tested as well as other classes of Arthropoda. Although we found no within‐species variation in this region, it provided enough information to separate closely related species or species complexes, in particular Thrips spp. and Bemisia spp. By combining a quick DNA extraction method with pyrosequencing, we were able to generate DNA sequences and complete species identification within 5 h. CONCLUSION: Mini‐barcode pyrosequencing of 16S rDNA coupled with the GenBank database provides a rapid, accurate, and efficient species identification system. This system is therefore useful for biodiversity discovery, forensic identification, and quarantine control and management. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Background: Amplification of small subunit (SSU) rRNA genes with universal primers is a common method used to assess microbial populations in various environmental samples. However, owing to limitations in coverage of these universal primers, some microorganisms remain unidentified. The present study aimed to establish a method for amplifying nearly full-length SSU rRNA gene sequences of previously unidentified prokaryotes, using newly designed targeted primers via primer evaluation in meta-transcriptomic datasets. Methods: Primer binding regions of universal primer 8F/Arch21F for bacteria or archaea were used for primer evaluation of SSU rRNA sequences in meta-transcriptomic datasets. Furthermore, targeted forward primers were designed based on SSU rRNA reads from unclassified groups unmatched with the universal primer 8F/Arch21F, and these primers were used to amplify nearly full-length special SSU rRNA gene sequences along with universal reverse primer 1492R. Similarity and phylogenetic analysis were used to confirm their novel status. Results: Using this method, we identified unclassified SSU rRNA sequences that were not matched with universal primer 8F and Arch21F. A new group within the Asgard superphylum was amplified by the newly designed specific primer based on these unclassified SSU rRNA sequences by using mudflat samples. Conclusion: We showed that using specific primers designed based on universal primer evaluation from meta-transcriptomic datasets, identification of novel taxonomic groups from a specific environment is possible. [ABSTRACT FROM AUTHOR]

Porcine astroviruses (PAstV) are members of the family Astroviridae , Mamastravirus genus and have been identified to have five genotypes (PAstV1–5). These viruses are highly prevalent in pigs and can cause enteric disease as well as neurological or respiratory symptoms depending on their genotypes. At present, the epidemiological impacts of some PAstV genotypes on pigs are largely unknown and hence continuously monitoring of these PAstVs may be needed. The purpose of this research was to develop an improved and efficient detection tool for PAstVs and to evaluate the developed method using clinical samples. Initially, a set of five chimeric primers (CP), each comprising genotype specific primer pairs with an identical universal adapter at the 5' end, and a universal primer (UP) that is identical to universal adapter sequence, were designed. With these tools in place, a novel multiplex PCR system with universal primer was established for the simultaneous detection of the five types of PAstV. This method can specifically detect PAstV genotypes, with a limit of detection (LOD) of 5 copies/μL for each genotype irrespective of single or mixed target template. Using this new assay, 273 pig fecal samples were investigated for further assay evaluation. Among all samples, the positive rate was 70.0% with PAstV4 in 56.8% of the samples, PAstV2 in 38.8%, PAstV1 in 16.8%, and PAstV5 in 11.0%. More than one PAstV in a sample were detected in 39.2% of the samples. The consistency rate between the novel multiplex PCR and singleplex PCRs was 96.4–100%. Given its rapidity, specificity and sensitivity, the novel multiplex PCR is a useful approach for demonstrating single or mixed genotype infections of PAstV. • A novel multiplex PAstV PCR was developed for detection and genotyping of PAstV. • This assay showed a high specificity and sensitivity with a detection limit of 5 copies per reaction. • This assay was proved to be a reliable diagnostic method for the detection and differentiation of five PAstV genotypes. [ABSTRACT FROM AUTHOR]

Copyright of Coatings & Protection / Tuceng yu Fanghu is the property of Coating & Protection Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Phytophthora infestans, the causal agent of late blight in potatoes and tomatoes, is the most important and ongoing pathogenic threat to agricultural production worldwide. Rapid and early identification of P. infestans is an essential prerequisite for countering the further spread of infection. In this study, a novel method for visual detection of P. infestans has been developed by integrating universal primer mediated asymmetric PCR with gold nanoparticle (AuNP)-based lateral flow biosensor. We employed asymmetric PCR to generate large amounts of single-stranded DNA (ssDNA) by amplifying a region of P. infestans -specific repetitive DNA sequence. The ssDNA products were then applied to the lateral flow biosensor to perform a visual detection using sandwich-type hybridization assays. In the presence of target DNA, sandwich-type hybridization reactions among the AuNP–probe, target DNA and capture probe were performed on the test line of the biosensor, and then a characteristic red band was produced for the accumulation of AuNPs. Quantitative analysis obtained by recording the optical intensity of the red band demonstrated that this biosensor could detect as little as 0.1 pg μL−1 genomic DNA. Furthermore, the specificity of the biosensor was confirmed by detecting three other Phytophthora species and two pathogenic fungi. We believe this method has potential application in early prediction of potato late blight disease and instigation of management actions to reduce the risk of epidemic development. Graphical abstract Image 1 Highlights • A gold nanoparticle-based lateral flow biosensor for visual detection of Phytophthora infestans has been developed. • The results of polyacrylamide gel electrophoresis confirmed the presence of the desired single-stranded DNA products. •; The specificity of the biosensor was confirmed by detecting three other Phytophthora species and two pathogenic fungi. • The biosensor was used to detect infected leaf and stem samples. [ABSTRACT FROM AUTHOR]

The present study aimed to develop a universal primer-multiplex PCR (UP-M-PCR) assay for the detection of six common bacteria associated with human meningitis. One optimal universal primer (UP) was selected from three UPs by comparing their sensitivities and specificities. All specific primers were tagged with the UP sequence at 5' end, and applied to the multiplex PCR system. The multiplex system was further optimized and assessed. This UP-M-PCR can successfully detect the six meningitis-associated pathogens with high specificity, and the sensitivity could reach up to 10 copies. In the identification of clinical specimens, six positive cases infected with Streptococcus agalactiae, Staphylococcus aureus, and Streptococcus pneumoniae were confirmed. The newly developed multiplex PCR system can be used to detect the six pathogens associated with human bacterial meningitis with high specificity and sensitivity. [ABSTRACT FROM AUTHOR]

We presented the powerful techniques for species identification using the short amplicon of mitochondrial cytochrome b gene sequence. Two faecal samples and one single hair sample of the Asian tapir were tested using the new cytochrome b primers. The results showed a high sequence similarity with the mainland Asian tapir group. The comparative sequence analysis of the reserved wild mammals in Thailand and the other endangered mammal species from Southeast Asia comprehensibly verified the potential of our novel primers. The forward and reverse primers were 94.2 and 93.2%, respectively, by the average value of the sequence identity among 77 species sequences, and the overall mean distance was 35.9%. This development technique could provide rapid, simple, and reliable tools for species confirmation. Especially, it could recognize the problematic biological specimens contained less DNA material from illegal products and assist with wildlife crime investigation of threatened species and related forensic casework. [ABSTRACT FROM AUTHOR]

Mycoplasmosis is one of the most important diseases in the poultry industry. Its causative agent, mycoplasma has various species, which two of them, Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most important species. Due to the enormous losses in the production farms of industrial poultry, achieving a rapid, accurate and definite diagnosis of mycoplasma is of great importance. An early and definite diagnosis can guarantee the farm management on keeping herd health. In many countries such as Iran, the disease and its complications have still remained as a serious problem. Given this issue, we decided to identify the mycoplasma infection from broiler poultry flocks through culture method. 150 carcasses of broiler chicken belonging to 50 broiler flocks were sampled in which the signs of air sacs involvement and secretions in the airways, trachea and bronchi were seen. Samples taken from trachea, palatine cleft, nasal passages and air sacs, were cultivated into PPLO liquid medium using membrane filters (0.45 micron). They were incubated at 37 °C and were examined for pH (color) changes for every 48 hours. During the first 24 hours after cultivation, every color change to yellow or dark visible with the bare eye was considered as bacterial contamination, therefore, the contaminated samples were removed from the incubator. The color change in the liquid media was compared with the uninoculated medium as negative control. If a color change was observed in the liquid media after 48h, subculture was done in the PPLO agar. The plates were incubated at 37 °C for 14 days. They were examined for mycoplasma colonies using a microscope with magnification of 10 in every other day. The results showed that out of 150 samples obtained from 50 broiler flocks, 16 (10.66%) were positive for mycoplasma, while in terms of contamination, 4 flocks (8%) were positive. The contamination of positive cultures was finally confirmed through PCR method with universal primer.

The purpose of this study was to develop a DNA marker for the identification of fish species in processed surimi products. A DNA marker was designed based on the mitochondrial cytochrome c oxidase subunit I gene, and fish species in surimi products were identified using a molecular fingerprinting technique, denaturing gradient gel electrophoresis (DGGE); the results were subjected to sequence-based analysis. The DGGE profiles indicated the presence in surimi products of a greater diversity of fish species than reported previously: 20 species belonging to 16 genera were identified. Therefore, our method facilitates the simple and rapid detection and identification of fish species in seafood products produced from minced fish. [ABSTRACT FROM AUTHOR]

There is an increasing requirement for traceability of aquaculture products, both for consumer protection and for food safety. There are high error rates in the conventional traceability systems depending on physical labels. Genetic traceability technique depending on DNA-based tracking system can overcome this problem. Genealogy information is essential for genetic traceability, and microsatellite DNA marker is a good choice for pedigree analysis. As increasing genotyping throughput of microsatellites, microsatellite multiplex PCR has become a fast and cost-effective technique. As a commercially important cultured aquatic species, Pacific oyster Crassostrea gigas has the highest global production. The objective of this study was to develop microsatellite multiplex PCR panels with dye-labeled universal primer for pedigree analysis in C. gigas, and these multiplex PCRs were validated using 12 full-sib families with known pedigrees. Here we developed six informative multiplex PCRs using 18 genomic microsatellites in C. gigas. Each multiplex panel contained a single universal primer M13(−21) used as a tail on each locus-specific forward primer and a single universal primer M13(−21) labeled with fluorophores. The polymorphisms of the markers were moderate, with an average of 10.3 alleles per locus and average polymorphic information content of 0.740. The observed heterozygosity per locus ranged from 0.492 to 0.822. Cervus simulations revealed that the six panels would still be of great value when massive families were analysed. Pedigree analysis of real offspring demonstrated that 100% of the offspring were unambiguously allocated to their parents when two multiplex PCRs were used. The six sets of multiplex PCRs can be an important tool for tracing cultured individuals, population genetic analysis, and selective breeding program in C. gigas. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the effects of different surface treatments and thermocycling on shear bond strength (SBS) be-tween resin cement and zirconia-reinforced lithium-silicate (ZLS) ceramic., Materials and Methods: 96 ZLS ceramic specimens were randomly allocated to four different surface treatment groups: etch and silane (ES), etch and universal primer (EUP), self-etching primer (SEP), and sandblasting and silane (SS). Stan-dardized composite cylinders were bonded to surface-treated ZLS ceramic, after which SBS was obtained either after 24-h water storage only or with an additional 5000 thermal cycles (TC), resulting in eight subgroups (n = 12). After evaluation of failure mode under a stereomicroscope, representative SEM images were acquired. To examine areal average surface roughness (Sa), additional ZLS specimens were prepared and randomly allocated to 3 groups: hydrofluoric acid etching, self-etching primer, and sandblasting (n = 10). Supplementary specimens were examined using field-emission scanning electron microscopy (FE-SEM) (n = 2) and atomic force microscopy (AFM) (n = 2) to investigate their surface topographies., Results: ANOVA showed a statistically significant difference in SBS following different surface treatment protocols after 24-h water storage (p < 0.001). However, TC groups revealed no statistically significant difference in their SBS (p = 0.394). All surface treated groups were significantly affected by TC (p < 0.001), except for the SS group (p = 0.48). Sa was signifi-cantly influenced by the different surface treatment protocols (p < 0.001)., Conclusion: The ability of self-etching primer to achieve comparable bond strength with a less technique-sensitive ap-proach makes it a favorable alternative to ES for the surface treatment of ZLS ceramics.

Gelatin, a purified protein derived mostly from pig skin and bovine tissue, is used widely in both food and pharmaceutical industries. Here, to determine the species of origin of capsule gelatin, we developed a sensitive and reliable test using the polymerase chain reaction (PCR) method, which included 1) species-specific or universal primer sets, designed to detect short 16S ribosomal RNA (rRNA) gene sequences from cow, pig, and fish (tilapia) as well as genes encoding the large subunit of plant ribulose-1,5-bisphosphate carboxylase oxygenase and 2) species-specific PCR coupled with whole-genome amplification. This method was used to verify manufacturing label claims of 28 gelatin capsule samples sold as dietary supplements. The results from 27 samples were consistent with gelatin-related information on the manufacturer label, while one sample that mentioned tilapia gelatin was found to contain only bovine DNA. This rapid method can therefore be used to verify the authenticity of gelatin capsules. [ABSTRACT FROM AUTHOR]

This study examines the potential of next-generation sequencing based ‘genotyping-by-sequencing’ (GBS) of microsatellite loci for rapid and cost-effective genotyping in large-scale population genetic studies. The recovery of individual genotypes from large sequence pools was achieved by PCR-incorporated combinatorial barcoding using universal primers. Three experimental conditions were employed to explore the possibility of using this approach with existing and novel multiplex marker panels and weighted amplicon mixture. The GBS approach was validated against microsatellite data generated by capillary electrophoresis. GBS allows access to the underlying nucleotide sequences that can reveal homoplasy, even in large datasets and facilitates cross laboratory transfer. GBS of microsatellites, using individual combinatorial barcoding, is potentially faster and cheaper than current microsatellite approaches and offers better and more data.

Using sequence alignment, a conserved domain in the 3' untranslated region (UTR) of the cytoplasmic heat shock protein 90 (HSP90) of Lepidoptera was found. This region is highly variable in other insect groups. Furthermore, universal primers were designed to amplify the complete coding sequence (CDS) of HSP90 from total genomic DNA in Lepidoptera, avoiding the commonly used reverse transcription-polymerase chain reaction (RT-PCR) and 3', 5'-rapid amplification of cDNA ends (RACE) methods based on cDNA. These primers amplified a fragment of about 2.25 kb in the 11 species tested, which represent seven different families of Lepidoptera, including moths and butterflies. The results suggest that the conserved domain of 3'UTR is universal in Lepidoptera and these primers successfully amplify the complete CDS of cytoplasmic HSP90 from genomic DNA.

Copyright of Journal of Orofacial Orthopedics/Fortschritte der Kieferorthopadie is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The aim of this study was to develop a cost-effective genotyping method using high-quality DNA for human identification. A total of 21 short tandem repeats (STRs) and amelogenin were selected, and fluorescent fragments at 22 loci were simultaneously amplified in a single-tube reaction using locus-specific primers with 24-base universal tails and four fluorescent universal primers. Several nucleotide substitutions in universal tails and fluorescent universal primers enabled the detection of specific fluorescent fragments from the 22 loci. Multiplex polymerase chain reaction (PCR) produced intense FAM-, VIC-, NED-, and PET-labeled fragments ranging from 90 to 400 bp, and these fragments were discriminated using standard capillary electrophoretic analysis. The selected 22 loci were also analyzed using two commercial kits (the AmpFLSTR Identifiler Kit and the PowerPlex ESX 17 System), and results for two loci (D19S433 and D16S539) were discordant between these kits due to mutations at the primer binding sites. All genotypes from the 100 samples were determined using 2.5 ng of DNA by our method, and the expected alleles were completely recovered. Multiplex 22-locus genotyping using four fluorescent universal primers effectively reduces the costs to less than 20% of genotyping using commercial kits, and our method would be useful to detect silent alleles from commercial kit analysis. [ABSTRACT FROM AUTHOR]

A novel multiplex PCR assay using non-homologous oligonucleotides with locked nucleic acid (LNA) modifications as universal primers was developed and validated for the simultaneous detection of five swine viruses. The assay utilizes five virus-specific primer pairs modified at the 5′ end through the addition of the universal primer sequence. In the reaction, small amounts of target templates with the 5′ tail were generated and subsequently amplified through the extension of a LNA universal primer set. To validate the specificity of this assay, 27 viral target strains and 12 non-target pathogens were tested. The lower limit of detection of viral nucleic acids was 1.1–1.9 pg per reaction or 11–32 pg in a five-plex viral nucleic acid mixture. The LNA mPCR assay displayed higher analytical sensitivity and efficiency for the detection of plasmid standards compared with the conventional assay, which uses standard primers without the 5′ tail. A total of 207 field samples were tested using both assays. The LNA mPCR assay provided numerically higher detection rates for all pathogens in independent samples. Moreover, the LNA mPCR assay had significantly higher detection rates in independent samples compared with the conventional assay. [ABSTRACT FROM AUTHOR]

Coagulase-positive and coagulase negative, methicillin-resistant staphylococci are major causes of serious nosocomial infections and it is very important to have a reliable test to detect these bacteria. A multiplex polymerase chain reaction (mPCR) was used on 100 clinical samples for simultaneous amplification of the universal bacterial, mec-A encoding the penicillin binding protein 2a, which is associated with staphylococcal methicillin resistance and TEM-1 encoding the β-lactamase, which accounts for the majority of all cases of the plasmid β-lactamase resistance worldwide. Out of 100 wound swabs tested, 99% with universal primers, 26% with TEM-1 primers and 6% with mec-A primers were positive. Dot blot Digoxigenin hybridization on the 30 samples was carried out to confirm identified bacteria with specific bacterial probes. Out of 100 wound swabs, 38% were positive with Staphylococcus aureus probe, 23% were positive with enteric bacteria probe, 7% were positive with Streptococcus agalactia probe and 1% were positive with Haemophilus influenza probe. The mPCR method used in this study, was designed to be incorporated into the workflow of the clinical microbiology laboratory and allows for the identification of intrinsic resistance in a timely and reliable manner.

In this study, we applied Non-dominated Sorting Genetic Algorithm II (NSGA-II) to the problem of identifying appropriate locations in cytochrome oxidase I (COI) gene for species- or group-specific primer design. As concerns about ecological management grow, quantification of predator-prey interaction has become a central issue in ecology. Recently, improved techniques have allowed for the extensive use of genomic DNA barcoding for qualitative analysis. However, quantification of DNA barcoding results is important and still needs to be resolved. Even though species- or group-specific primers that can be used for samples containing multiple species are difficult to design, species- or group-specific primers are a practical solution for quantification in the current era. To resolve this issue, we present here an efficient method for discovering the regions of a DNA sequence that have the highest inter-species variability by applying the NSGA-II algorithm. DNA sequence information for the COI gene region was obtained for 24 species from Jo et al. (2014). These sequences were transformed into binary data, either 0 (not different) or 1 (different), to reflect sequence conservation at each base for all combinations of two species. These data were analyzed by two objective functions, the average and standard deviation of the difference, which were used in the NSGA-II algorithm to search for appropriate DNA locations for species-specific primer design. The NSGA-II program identified four solutions (possible primer binding sites); consequently NSGA-II is believed to be a suitable algorithm for species-specific primer design, and is expected to make this difficult and time-consuming process more efficient. [ABSTRACT FROM AUTHOR]

Purpose: To detect Strongyloides ratti in faecal samples using conventional methods and to confirm the identification using a sensitive and specific method, namely, polymerase chain reaction (PCR). Methods: A PCR method targeting the small subunit of the rRNA gene was performed in this study for the detection of DNA from Strongyloides ratti (an animal model of S. stercoralis) in faecal samples of wild Brown rats, Rattus norvegicus. Results: Strongyloides ratti was detected in 34.2 % of collected rats by different conventional techniques and confirmed by PCR. The essay presented 100 % sensitivity with Strongyloides universal primer. Conclusion: The findings of this study suggest that the application of PCR with universal primer is a very sensitive methodology to detect S. ratti in faecal material of wild rats infected even with very low parasite burden. [ABSTRACT FROM AUTHOR]

Amplification of fluorescently labeled products is one of the most popular methods for genotyping genetic variations. Two-step amplification using fluorescent universal primers simultaneously produces multiple targeted fragments labeled with fluorescent dyes, and this strategy is applicable to large-scale, cost-effective genotyping. In this study, we developed a fast PCR-based, multiple short tandem repeat (STR) genotyping method using fluorescent universal primers containing locked nucleic acids (LNAs). Four amplification reactions, each assaying six or seven markers and using 0.5–1.0 ng of genomic DNA, produced obvious Fam-labeled peaks in all 26 loci tested (25 autosomal STRs and amelogenin). The overall amplification time was 37 min. Moreover, fluorescent signals for the 25 STRs obtained from LNA-containing primers were 1.5–9.0 fold higher compared to those from non-LNA primers. Using genomic DNA from 120 Japanese individuals, 16 out of the 25 STRs had observed heterozygosity greater than 0.7. Some of these 25 STRs also had high discriminatory power, similar to that of the 13 core STRs in the Combined DNA Index System dataset. The probability of incorrectly assigning a match based on the accumulated matching probability for these 25 STRs is 1.2 × 10 −22 , and their combined use can provide robust information for Japanese forensics. [ABSTRACT FROM AUTHOR]

A number of viruses from the genera Furovirus, Hordeivirus and Rymovirus are known to infect and damage the four major temperate cereal crops, wheat, barley, sorghum and oats. Currently, there is no active testing in Australia for any of these viruses, which pose a significant biosecurity threat to the phytosanitary status of Australia's grains industry. To address this, broad spectrum PCR assays were developed to target virus species within the genera Furovirus, Hordeivirus and Rymovirus . Five sets of novel genus-specific primers were designed and tested in reverse-transcription polymerase chain reaction assays against a range of virus isolates in plant virus diagnostic laboratories in both Australia and New Zealand. Three of these assays were then chosen to screen samples in a three-year survey of cereal crops in western Victoria, Australia. Of the 8900 cereal plants screened in the survey, all were tested free of furoviruses, hordeiviruses and rymoviruses. To date, there were no published genus-specific primers available for the detection of furoviruses, hordeiviruses and rymoviruses. This study shows for the first time a broad-spectrum molecular test being used in a survey for exotic grain viruses in Australia. Results from this survey provide important evidence of the use of this method to demonstrate the absence of these viruses in Victoria, Australia. The primer pairs reported here are expected to detect a wide range of virus species within the three genera. [ABSTRACT FROM AUTHOR]

Purpose: This study will evaluate the effectiveness of various pretreatments when fiber-reinforced composite (FRC) post is bonded to endodontically treated tooth with resin cement. Materials and methods: Canal shaping of FRC post (DT Light post, Size 3, Bisco Inc., Schaumburg, IL, USA) was performed on endodontically treated premolars at 1.5 cm from CEJ. Samples were divided into 6 groups of surface treatment after conventional washing and drying to the canal. Total of 24 FRC posts were randomly divided into 6 groups of surface treatment as follows: Group C: control - no surface treatment, Group A: airborne-particle abrasion (Cojet sand, 3M ESPE), Group S: silanization (Bis-silane, Bisco Inc.), Group M: universal primer (Monobond-plus primer, Ivoclar Vivadent Inc.), Group AS: silanization after airborne-particle abrasion, Group AM: universal primer treatment after airborne-particle abrasion. Pretreated fiber posts were cemented with resin-based luting material and photo-polymerized and cut to the thickness of 1 mm. Push-out test using a universal testing machine was performed. Bonding failure strength of post dislodgement was measured and the type of bonding failure was classified. Data were analyzed with Kruskal-Wallis test and multiple comparison groups were performed using Tukey HSD value of rank test (a=0.05). Results: Group AS showed significantly highest bonding strength. Group S, group AM, group A, and group M showed lower bonding strength in order. The control group showed the lowest bonding strength. Conclusion: Surface treatment with silane showed to be the most effective of the surface pretreatment methods for cementation of FRC post. Surface treatment with universal primer showed no significant difference compared with no surface treatment group as for bonding strength. [ABSTRACT FROM AUTHOR]