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Rapid and sensitive detection of Salmonella with reduced graphene oxide-carbon nanotube based electrochemical aptasensor

Appaturi JN, Pulingam T, Thong KL, Muniandy S, Ahmad N, Leo BF

Anal Biochem, 2020 01 15;589:113489.
PMID: 31655050 DOI: 10.1016/j.ab.2019.113489

Rapid detection of foodborne pathogens is crucial as ingestion of contaminated food products may endanger human health. Thus, the objective of this study was to develop a biosensor using reduced graphene oxide-carbon nanotubes (rGO-CNT) nanocomposite via the hydrothermal method for accurate and rapid label-free electrochemical detection of pathogenic bacteria such as Salmonella enterica. The rGO-CNT nanocomposite was characterized using Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray diffraction and transmission electron microscopy. The nanocomposite was dropped cast on the glassy carbon electrode and further modified with amino-modified DNA aptamer. The resultant ssDNA/rGO-CNT/GCE aptasensor was then used to detect bacteria by using differential pulse voltammetry (DPV) technique. Synergistic effects of aptasensor was evident through the combination of enhanced electrical properties and facile chemical functionality of both rGO and CNT for the stable interface. Under optimal experimental conditions, the aptasensor could detect S. Typhimurium in a wide linear dynamic range from 101 until 108 cfu mL-1 with a 101 cfu mL-1 of the limit of detection. This aptasensor also showed good sensitivity, selectivity and specificity for the detection of microorganisms. Furthermore, we have successfully applied the aptasensor for S. Typhimurium detection in real food samples.

Matched MeSH terms: Limit of Detection
Divalent ion-induced aggregation of gold nanoparticles for voltammetry Immunosensing: comparison of transducer signals in an assay for the squamous cell carcinoma antigen

Letchumanan I, Gopinath SCB, Arshad MKM

Mikrochim Acta, 2020 01 14;187(2):128.
PMID: 31938893 DOI: 10.1007/s00604-020-4115-0

A method is described for the electrochemical determination of squamous cell carcinoma (SCC) antigen, and by testing the effect of 30 nm gold nanoparticles (GNPs). Three comparative studies were performed in the presence and absence of GNPs, and with agglomerated GNPs. The divalent ion Ca(II) was used to induce a strong agglomeration of GNPs, as confirmed by colorimetry and voltammetry. Herein, colorimetry was used to test the best amount of salt needed to aggregate the GNPs. Despite, voltammetry was used to determine the status of biomolecules on the sensor. The topography of the surface of ZnO-coated interdigitated electrodes was analyzed by using 3D-nano profilometry, scanning electron microscopy, atomic force microscopy and high-power microscopy. The interaction between SCC antigen and antibody trigger vibrations on the sensor and cause dipole moment, which was measured using a picoammeter with a linear sweep from 0 to 2 V at 0.01 V step voltage. The sensitivity level was 10 fM by 3σ calculation for the dispersed GNP-conjugated antigen. This indicates a 100-fold enhancement compared to the condition without GNP conjugation. However, the sensitivity level for agglomerated GNPs conjugated antibody was not significant with 100 fM sensitivity. Specificity was tested for other proteins in serum, namely blood clotting factor IX, C-reactive protein, and serum albumin. The SCC antigen was quantified in spiked serum and gave recoveries that ranged between 80 and 90%. Graphical abstractSchematic representation of SCC (squamous cell carcinoma) antigen determination using divalent ion induced agglomerated GNPs. Sensitivity increment depends on the occurrence of more SCC antigen and antibody binding event via GNPs integration. Notably, lower detection limit was achieved at femto molar with proper orientation of biological molecules.

Matched MeSH terms: Limit of Detection
Fulltext Early-stage cervical cancer diagnosis based on an ultra-sensitive electrochemical DNA nanobiosensor for HPV-18 detection in real samples

Mahmoodi P, Rezayi M, Rasouli E, Avan A, Gholami M, Ghayour Mobarhan M, et al.

J Nanobiotechnology, 2020 Jan 13;18(1):11.
PMID: 31931815 DOI: 10.1186/s12951-020-0577-9

BACKGROUND: In several years ago, infection with human papillomaviruses (HPVs), have been prevalent in the worlds especially HPV type 18, can lead to cervical cancer. Therefore, rapid, accurate, and early diagnosis of HPV for successful treatment is essential. The present study describes the development of a selective and sensitive electrochemical biosensor base on DNA, for early detection of HPV-18. For this purpose, a nanocomposite of reduced graphene oxide (rGO) and multiwalled carbon nanotubes (MWCNTs) were electrodeposited on a screen-printed carbon electrode (SPCE). Then, Au nanoparticles (AuNPs) were dropped on a modified SPCE. Subsequently, single strand DNA (ssDNA) probe was immobilized on the modified electrode. The link attached between AuNPs and probe ssDNA provided by L-cysteine via functionalizing AuNPs (Cys-AuNPs). The differential pulse voltammetry (DPV) assay was also used to electrochemical measurement. The measurement was based on the oxidation signals of anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) before and after hybridization between the probe and target DNA.
RESULTS: The calibration curve showed a linear range between 0.01 fM to 0.01 nM with a limit of detection 0.05 fM. The results showed that the optimum concentration for DNA probe was 5 µM. The good performance of the proposed biosensor was achieved through hybridization of DNA probe-modified SPCE with extracted DNA from clinical samples.
CONCLUSIONS: According to the investigated results, this biosensor can be introduced as a proprietary, accurate, sensitive, and rapid diagnostic method of HPV 18 in the polymerase chain reaction (PCR) of real samples.

Matched MeSH terms: Limit of Detection
Using SERS-based microfluidic paper-based device (μPAD) for calibration-free quantitative measurement of AMI cardiac biomarkers

Lim WY, Goh CH, Thevarajah TM, Goh BT, Khor SM

Biosens Bioelectron, 2020 Jan 01;147:111792.
PMID: 31678828 DOI: 10.1016/j.bios.2019.111792

Recently, surface enhanced Raman scattering (SERS) has attracted much attention in medical diagnosis applications owing to better detection sensitivity and lower limit of detection (LOD) than colorimetric detection. In this paper, a novel calibration-free SERS-based μPAD with multi-reaction zones for simultaneous quantitative detection of multiple cardiac biomarkers - GPBB, CK-MB and cTnT for early diagnosis and prognosis of acute myocardial infarction (AMI) are presented. Three distinct Raman probes were synthesised, subsequently conjugated with respective detecting antibodies and used as SERS nanotags for cardiac biomarker detection. Using a conventional calibration curve, quantitative simultaneous measurement of multiple cardiac biomarkers on SERS-based μPAD was performed based on the characteristic Raman spectral features of each reporter used in different nanotags. However, a calibration free point-of-care testing device is required for fast screening to rule-in and rule-out AMI patients. Partial least squares predictive models were developed and incorporated into the immunosensing system, to accurately quantify the three unknown cardiac biomarkers levels in serum based on the previously obtained Raman spectral data. This method allows absolute quantitative measurement when conventional calibration curve fails to provide accurate estimation of cardiac biomarkers, especially at low and high concentration ranges. Under an optimised condition, the LOD of our SERS-based μPAD was identified at 8, 10, and 1 pg mL-1, for GPBB, CK-MB and cTnT, respectively, which is well below the clinical cutoff values. Therefore, this proof-of-concept technique shows significant potential for highly sensitive quantitative detection of multiplex cardiac biomarkers in human serum to expedite medical decisions for enhanced patient care.

Matched MeSH terms: Limit of Detection
Fulltext Evaluation of extra virgin olive oil adulteration with edible oils using ATR-FTIR spectroscopy

Nuraznee Mashodi, Nurul Yani Rahim, Norhayati Muhammad, Saliza Asman

Malaysian Journal of Applied Sciences, 2020;5(1):35-44.
MyJurnal

Extra virgin olive oil (EVOO) is categorized as expensive oil due to high-quality nutritional value. Unfortunately, EVOO is easily adulterated with other low-quality edible oils. Therefore, this study was done to differentiate and analyze the adulteration of EVOO with other edible oils using Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The study was used several edible oils included canola oil, corn oil, sunflower oil, and soybean oil as an adulterant for EVOO. The adulterant EVOO samples were prepared by mixing with dissimilar concentrations of the solely edible oils (20 %, 40 %, 60 % and 80 % (v/v)). The main functional groups of EVOO and other edible oils are O-H, C-H, C=C and C=O groups were assigned around 3500 cm-1, 2925 cm-1, 3006 cm-1 and 1745 cm-1 wavenumbers, respectively. From the comparison of EVOO and other adulterant edibles oil spectra, it showed that the EVOO has the lowest absorbance intensity at around 3006 cm-1 represented double bond which is closely related to the composition of oil sample. The adulteration of EVOO was evaluated by analysing the changes in the absorbance based on the linear regression analysis graph of the bands at 3006 and 2925 cm-1 and the limit of detection (LOD) was measured. The graph of A3008/A2925 with good relative coefficients (R2) and lower LOD is more favourable than the linear regression graph of A3006 versus percentage of edible oils added in EVOO. This study showed that ATR-FTIR spectroscopy is a convenient tool for analysing the adulteration of EVOO.

Matched MeSH terms: Limit of Detection
Fulltext Rapid identification of melioidosis agent by an insulated isothermal PCR on a field-deployable device

Chua KH, Tan EW, Chai HC, Puthucheary SD, Lee PC, Puah SM

PeerJ, 2020;8:e9238.
PMID: 32518734 DOI: 10.7717/peerj.9238

Background: Burkholderia pseudomallei causes melioidosis, a serious illness that can be fatal if untreated or misdiagnosed. Culture from clinical specimens remains the gold standard but has low diagnostic sensitivity.
Method: In this study, we developed a rapid, sensitive and specific insulated isothermal Polymerase Chain Reaction (iiPCR) targeting bimA gene (Burkholderia Intracellular Motility A; BPSS1492) for the identification of B. pseudomallei. A pair of novel primers: BimA(F) and BimA(R) together with a probe were designed and 121 clinical B. pseudomallei strains obtained from numerous clinical sources and 10 ATCC non-targeted strains were tested with iiPCR and qPCR in parallel.
Results: All 121 B. pseudomallei isolates were positive for qPCR while 118 isolates were positive for iiPCR, demonstrating satisfactory agreement (97.71%; 95% CI [93.45-99.53%]; k = 0.87). Sensitivity of the bimA iiPCR/POCKIT assay was 97.52% with the lower detection limit of 14 ng/µL of B. pseudomallei DNA. The developed iiPCR assay did not cross-react with 10 types of non-targeted strains, indicating good specificity.
Conclusion: This bimA iiPCR/POCKIT assay will undoubtedly complement other methodologies used in the clinical laboratory for the rapid identification of this pathogen.

Matched MeSH terms: Limit of Detection
Ultrasonic synthesis of CuO nanoflakes: A robust electrochemical scaffold for the sensitive detection of phenolic hazard in water and pharmaceutical samples

Hwa KY, Karuppaiah P, Gowthaman NSK, Balakumar V, Shankar S, Lim HN

Ultrason Sonochem, 2019 Nov;58:104649.
PMID: 31450344 DOI: 10.1016/j.ultsonch.2019.104649

Hydroquinone (HQ), a phenolic compound is expansively used in many industrial applications and due to the utilization of HQ, water pollution tragedies frequently found by the improper handling and accidental outflows. When HQ is adsorbed directly through the skin that create toxic effects to human by affecting kidney, liver, lungs, and urinary tract and hence, a highly selective and sensitive technique is required for its quantification. Herein, we have developed the ultrasonic synthesis of copper oxide nanoflakes (CuO-NFs) using ultrasonic bath (20 kHz, 100 W) and successfully employed for the sensitive detection of the environmental hazardous pollutant HQ. The formed CuO-NFs were confirmed by X-ray diffraction, field emission scanning electron microscopy (FE-SEM), FT-IR spectroscopy and UV-visible spectroscopy and fabricated with the screen-printed carbon electrode (SPCE). The SEM images exhibited the uniform CuO-NFs with an average width of 85 nm. The linker-free CuO-NFs fabricated electrode showed the appropriate wide range of concentrations from 0.1 to 1400 µM and the limit of detection was found to be 10.4 nM towards HQ. The fabricated sensor having long term stability and sensitivity was successfully applied for the environmental and commercial real sample analysis and exhibited good recovery percentage, implying that the SPCE/CuO-NFs is an economically viable and benign robust scaffold for the determination of HQ.

Matched MeSH terms: Limit of Detection*
Fabricated germanium-doped fibres for computed tomography dosimetry

Rais NNM, Bradley DA, Hashim A, Osman ND, Noor NM

Appl Radiat Isot, 2019 Nov;153:108810.
PMID: 31351374 DOI: 10.1016/j.apradiso.2019.108810

For a range of doses familiarly incurred in computed tomography (CT), study is made of the performance of Germanium (Ge)-doped fibre dosimeters formed into cylindrical and flat shapes. Indigenously fabricated 2.3 mol% and 6 mol% Ge-dopant concentration preforms have been used to produce flat- and cylindrical-fibres (FF and CF) of various size and diameters; an additional 4 mol% Ge-doped commercial fibre with a core diameter of 50 μm has also been used. The key characteristics examined include the linearity index f(d), dose sensitivity and minimum detectable dose (MDD), the performance of the fibres being compared against that of lithium-fluoride based TLD-100 thermoluminescence (TL) dosimeters. For doses in the range 2-40 milligray (mGy), delivered at constant potential of 120 kilovoltage (kV), both the fabricated and commercial fibres demonstrate supralinear behaviours at doses  4 mGy. In terms of dose sensitivity, all of the fibres show superior TL sensitivity when compared against TLD-100, the 2.3 mol% and 6 mol% Ge-doped FF demonstrating the greatest TL sensitivity at 84 and 87 times that of TLD-100. The TL yields for the novel Ge-doped silica glass render them appealing for use within the present medical imaging dose range, offering linearity at high sensitivity down to less than 2 mGy.

Matched MeSH terms: Limit of Detection
Determination of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes using liquid chromatography-high-resolution mass spectrometry (LC-HRMS)

Mohd Yusop AY, Xiao L, Fu S

Talanta, 2019 Nov 01;204:36-43.
PMID: 31357306 DOI: 10.1016/j.talanta.2019.05.078

As a widely consumed beverage, coffee tends to be a target for intentional adulteration. This study describes the application of modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) coupled to liquid chromatography-high-resolution mass spectrometry (LC-HRMS) for simultaneous screening, identification, and quantification of undeclared phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes (ICPs). The mass spectrometer was operated in auto MS/MS acquisition for simultaneous MS and MS/MS experiments. Qualitative establishments from the suspected-target screening and targeted identification processes led to an unambiguous analyte assignment from the protonated molecule ([M+H]+) precursor ion which is subsequently used for quantification of 23 targeted PDE5 inhibitors. The analytical method validation covered specificity, linearity, range, accuracy, limit of detection (LOD), limit of quantification (LOQ), precisions, matrix effect (ME), and extraction recovery (RE). The specificity was established using the optimised chromatographic separation as well as the distinguishable [M+H]+ precursor ion. The linearity of each target analyte was demonstrated with a coefficient of determination (r2) of >0.9960 over the expected range of sample concentrations. The accuracy ranged from 88.1%-119.3% with LOD and LOQ of <70 ng/mL and 80 ng/mL, respectively. Excellent precisions were established within 0.4%-9.1% of the relative standard deviation. An insignificant ME within -5.2% to +8.7% was achieved using three different strategies of chromatography, sample extraction, and sample dilution. The RE was good for all target analytes within 84.7%-123.5% except for N-desethylacetildenafil at low (53.8%) and medium (65.1%) quality control levels. The method was successfully applied to 25 samples of ICPs where 17 of them were found to be adulterated with PDE5 inhibitors and their analogues. Further quantification revealed the total amount of these adulterants ranged from 2.77 to 121.64 mg per sachet.

Matched MeSH terms: Limit of Detection
Electrocatalytic Reduction of Hydrogen Peroxide and Its Non-Enzymatic Electrochemical Detection Using Silver Nanoparticles Anchored on Reduced Graphene Oxide

Ikhsan NI, Rameshkumar P, Yusoff N, Huang NM

J Nanosci Nanotechnol, 2019 Nov 01;19(11):7054-7063.
PMID: 31039858 DOI: 10.1166/jnn.2019.16630

Silver-reduced graphene oxide (Ag-rGO) nanohybrid was synthesized by applying a slight modification to the Turkevich method using trisodium citrate as a reducing and stabilizing agent to catalyze the non-enzymatic electrochemical detection of hydrogen peroxide (H₂O₂). Spherical silver nanoparticles (AgNPs) with an average particle size of 2.2 nm surfaced on reduced graphene oxide (rGO) sheets. Cyclic voltammograms (CV) obtained from glassy carbon (GC) electrode coated with Ag-rGO nanohybrid (4 mM) exhibited a peak at an overpotential of -0.52 V, with a larger faradaic current for the reduction of H₂O₂. Using the modified electrode for the linear sweep voltammetry (LSV) detection of H₂O₂, the detection limit and sensitivity were determined to be 4.8 μM (S/N ═ 3) and 0.0262 μA μM-1, respectively. The sensor appeared selective and stable towards H₂O₂ in the presence of possible interference, and it also demonstrated good recoveries of H₂O₂ concentration in real water samples.

Matched MeSH terms: Limit of Detection
Fulltext Single-tube, dual channel pentaplexing for the identification of Candida strains associated with human infection

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: Limit of Detection
Fulltext Subtractive inhibition assay for the detection of Campylobacter jejuni in chicken samples using surface plasmon resonance

Masdor NA, Altintas Z, Shukor MY, Tothill IE

Sci Rep, 2019 09 20;9(1):13642.
PMID: 31541137 DOI: 10.1038/s41598-019-49672-2

In this work, a subtractive inhibition assay (SIA) based on surface plasmon resonance (SPR) for the rapid detection of Campylobacter jejuni was developed. For this, rabbit polyclonal antibody with specificity to C. jejuni was first mixed with C. jejuni cells and unbound antibody was subsequently separated using a sequential process of centrifugation and then detected using an immobilized goat anti-rabbit IgG polyclonal antibody on the SPR sensor chip. This SIA-SPR method showed excellent sensitivity for C. jejuni with a limit of detection (LOD) of 131 ± 4 CFU mL-1 and a 95% confidence interval from 122 to 140 CFU mL-1. The method has also high specificity. The developed method showed low cross-reactivity to bacterial pathogens such as Salmonella enterica serovar Typhimurium (7.8%), Listeria monocytogenes (3.88%) and Escherichia coli (1.56%). The SIA-SPR method together with the culturing (plating) method was able to detect C. jejuni in the real chicken sample at less than 500 CFU mL-1, the minimum infectious dose for C. jejuni while a commercial ELISA kit was unable to detect the bacterium. Since the currently available detection tools rely on culturing methods, which take more than 48 hours to detect the bacterium, the developed method in this work has the potential to be a rapid and sensitive detection method for C. jejuni.

Matched MeSH terms: Limit of Detection
An Electrochemical Sandwich Immunosensor for the Detection of HER2 using Antibody-Conjugated PbS Quantum Dot as a label

Lah ZMANH, Ahmad SAA, Zaini MS, Kamarudin MA

J Pharm Biomed Anal, 2019 Sep 10;174:608-617.
PMID: 31265987 DOI: 10.1016/j.jpba.2019.06.024

A facile electrochemical sandwich immunosensor for the detection of a breast cancer biomarker, the human epidermal growth factor receptor 2 (HER2), was designed, using lead sulfide quantum dots-conjugated secondary HER2 antibody (Ab2-PbS QDs) as a label. Using Ab2-PbS QDs in the development of electrochemical immunoassays leads to many advantages such as straightforward synthesis and well-defined stripping signal of Pb(II) through acid dissolution, which in turn yields better sensing performance for the sandwiched immunosensor. In the bioconjugation of PbS QDs, the available amine and hydroxyl groups from secondary anti-HER2 and capped PbS QDs were bound covalently together via carbonyldiimidazole (CDI) acting as a linker. In order to quantify the biomarker, SWV signal was obtained, where the Pb2+ ions after acid dissolution in HCl was detected. The plated mercury film SPCE was also detected in situ. Under optimal conditions, HER2 was detected in a linear range from 1-100 ng/mL with a limit of detection of 0.28 ng/mL. The measures of satisfactory recoveries were 91.3% to 104.3% for the spiked samples, displaying high selectivity. Therefore, this method can be applied to determine HER2 in human serum.

Matched MeSH terms: Limit of Detection
Zinc(II) salphen complex-based fluorescence optical sensor for biogenic amine detection

Sahudin MA, Su'ait MS, Tan LL, Lee YH, Abd Karim NH

Anal Bioanal Chem, 2019 Sep;411(24):6449-6461.
PMID: 31392436 DOI: 10.1007/s00216-019-02025-4

Biogenic amines have attracted interest among researchers because of their importance as biomarkers in determining the quality of food freshness in the food industry. A rapid and simple technique that is able to detect biogenic amines is needed. In this work, a new optical sensing material for one of the biogenic amines, histamine, based on a new zinc(II) salphen complex was developed. The binding of zinc(II) complexes without an electron-withdrawing group (complex 1) and with electron-withdrawing groups (F, complex 2; Cl, complex 3) to histamine resulted in enhancement of fluorescence. All complexes exhibited high affinity for histamine [binding constant of (7.14 ± 0.80) × 104, (3.33 ± 0.03) × 105, and (2.35 ± 0.14) × 105 M-1, respectively]. Complex 2 was chosen as the sensing material for further development of an optical sensor for biogenic amines in the following step since it displayed enhanced optical properties in comparison with complexes 1 and 3. The optical sensor for biogenic amines used silica microparticles as the immobilisation support and histamine as the analyte. The optical sensor had a limit of detection for histamine of 4.4 × 10-12 M, with a linear working range between 1.0 × 10-11 and 1.0 × 10-6 M (R2 = 0.9844). The sensor showed good reproducibility, with a low relative standard deviation (5.5 %). In addition, the sensor exhibited good selectivity towards histamine and cadaverine over other amines, such as 1,2-phenylenediamine, triethylamine, and trimethylamine. Recovery and real sample studies suggested that complex 2 could be a promising biogenic amine optical sensing material that can be applied in the food industry, especially in controlling the safety of food for it to remain fresh and healthy for consumption.

Matched MeSH terms: Limit of Detection
Fulltext The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples

Aina GQ, Erwanto Y, Hossain M, Johan MR, Ali ME, Rohman A

J Adv Vet Anim Res, 2019 Sep;6(3):300-307.
PMID: 31583226 DOI: 10.5455/javar.2019.f348

Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) in meatball products.
Materials and Methods: The novel SSP of CYTBWB2-wb was designed by our group using PRIMERQUEST and NCBI software. DNA was extracted using propanol-chloroform-isoamyl alcohol method. The designed SSP was further subjected for validation protocols using DNA isolated from fresh meat and from meatball, which include specificity test, determination of efficiency, limit of detection and repeatability, and application of developed method for analysis of commercially meatball samples.
Results: The results showed that CYTBWB2-wb was specific to wild boar species against other animal species with optimized annealing temperature of 59°C. The efficiency of q-PCR obtained was 91.9% which is acceptable according to the Codex Allimentarius Commission (2010). DNA, with as low as 5 pg/μl, could be detected using q-PCR with primer of CYTBWB2-wb. The developed method was also used for DNA analysis extracted from meatball samples commercially available.
Conclusion: q-PCR using CYTBWB2-wb primers targeting on mitochondrial cytochrome-b gene (forward: CGG TTC CCT CTT AGG CAT TT; Reverse: GGA TGA ACA GGC AGA TGA AGA) can be fruitfully used for the analysis of WBM in commercial meatball samples.

Matched MeSH terms: Limit of Detection
Detection and differentiation of opportunistic viral infections potentially contributing to renal graft rejection by tetraplex-nested PCR

Ashouri Saheli Z, Shenagari M, Harzandi N, Monfared A

Trop Biomed, 2019 Sep 01;36(3):766-775.
PMID: 33597498

The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EBV as compared with commercial kits. Regarding the sensitivity and specificity, it seems that the developed Multiplex Nested PCR assay could be used as a reliable virusassociated renal rejection (VRR) panel in post renal transplant surveillance.

Matched MeSH terms: Limit of Detection
Bioinspired 2D carbon sheets decorated with MnFe2O4 nanoparticles for preconcentration of inorganic arsenic, and its determination by ICP-OES

Ahmad H, Haseen U, Umar K, Ansari MS, Ibrahim MNM

Mikrochim Acta, 2019 08 27;186(9):649.
PMID: 31456042 DOI: 10.1007/s00604-019-3753-6

The authors describe a method for solvent-free mechano-chemical synthesis of a bioinspired sorbent. A 2D ultra-thin carbon sheet similar to graphene oxide was prepared using a natural waste (onion sheet). The formation of 2D carbon sheets was confirmed by Raman spectroscopy, X-ray photoelectron spectroscopy and ATR-IR. The surface morphology was characterized by field emission scanning electron microscopy and high-resolution tunneling electron microscopy. The carbon sheets were decorated with crystalline MnFe2O4 nanoparticles by solid-state reaction at room temperature. The presence of magnetic particles in the final product was confirmed by vibrating sample magnetometry and electron microscopy. The synergistic effect of carbon sheets and MnFe2O4 led to an enhanced sorption of arsenic species compared to bare carbon sheets or to MnFe2O4 nanoparticles. A column was prepared for the simultaneous preconcentration and determination of trace levels of As(III) and As(V) from water samples. The preconcentration factors are between 900 and 833 for As(III) and As(V) species, respectively. The linearity of the calibration plot ranges from 0.4-10 ng mL-1. The detection limits (at 3σ) for both As(III) and As(V) are 30 pg mL-1. The Student's t values for the analysis of spiked samples are lower than the critical Student's t values at a 95% confidence level. The recoveries from spiked water samples range between 99 and 102.8%. Graphical abstract Schematic representation of the preparation of carbon sheets similar to graphene oxide from onion sheaths after pyrolysis at 800 °C. The prepared carbon sheet-MnFe2O4 composite shows excellent arsenic sorption and preconcentration down to the pg mL-1 concentration.

Matched MeSH terms: Limit of Detection
Fulltext Salting-out assisted liquid-liquid extraction coupled with high-performance liquid chromatography for the determination of vitamin D3 in milk samples

Sazali NH, Alshishani A, Saad B, Chew KY, Chong MM, Miskam M

R Soc Open Sci, 2019 Aug;6(8):190952.
PMID: 31598260 DOI: 10.1098/rsos.190952

In this study, salting-out assisted liquid-liquid extraction (SALLE) as a simple and efficient extraction technique followed by high-performance liquid chromatography (HPLC) was employed for the determination of vitamin D3 in milk samples. The sample treatment is based on the use of water-miscible acetonitrile as the extractant and acetonitrile phase separation under high-salt conditions. Under the optimum conditions, acetonitrile and ammonium sulfate were used as the extraction solvent and salting-out agent, respectively. The vitamin D3 extract was separated using Hypersil ODS (250x i.d 4.6 mm, 5 µm) HPLC column that was coupled with diode array detector. Vitamin D2 was used as internal standard (IS) to offset any variations in chromatographic conditions. The vitamin D3 and the IS were eluted in 18 min. Good linearity (r2 > 0.99) was obtained within the range of 25-600 ng g-1 with the limit of detection of 15 ng g-1 and limit of quantification of 25 ng g-1. The validated method was applied for the determination of vitamin D3 in milk samples. The recoveries for spiked samples were from 94.4 to 113.5%.

Matched MeSH terms: Limit of Detection
Voltammetric determination of iodide in iodized table salt using cetyltrimethylammonium bromide as ion-pairing

Jamilan MA, Abdullah J, Alang Ahmad SA, Md Noh MF

J Food Sci Technol, 2019 Aug;56(8):3846-3853.
PMID: 31413410 DOI: 10.1007/s13197-019-03855-x

In this work, voltammetric study based on cetyltrimethylammonium bromide (CTAB) as an ion-pairing agent for the determination of iodine level in iodized table salt has been explored. CTAB was used as an intermediate compound between iodide (I-) and the electrode due to its ability to dissociate to produce cetyltrimethylammonium ions ([CTA]+). The [CTA]+ with a long hydrophobic alkyl chain can be directly adsorbed onto the surface of the working electrode, and this in turns coated the electrode with cationic charge and enhance the electrode ability to bind to iodide (I-) and other molecular iodine ions. A mixture of iodide and CTAB ([CTA]+I-) was prepared and potential of 1.0 V for 60.0 s was applied to pre-concentrate the solution on the working electrode causing the [CTA]+I- to oxidize to iodine (I2). The produced I2 immediately react with chloride ion (Cl-) from the electrolyte of hydrochloric acid (HCl) to produce I2Cl- and form ion-pair with CTA+ as [CTA]+I2Cl-. The linear calibration curve of the developed method towards iodide was in the concentration range of 0.5-4.0 mg/L with sensitivity of - 1.383 µA mg/L-1 cm-2 (R2 = 0.9950), limit of detection (LOD) of 0.3 mg/L and limit of quantification (LOQ) of 1.0 mg/L, respectively. The proposed method indicates good agreement with the standard method for iodine determination with recovery range from 95.0 to 104.3%. The developed method provided potential application as a portable on-site iodine detector.

Matched MeSH terms: Limit of Detection
Fulltext Data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes

Mohd Yusop AY, Xiao L, Fu S

Data Brief, 2019 Aug;25:104234.
PMID: 31384643 DOI: 10.1016/j.dib.2019.104234

This paper presents the data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors and their analogues as adulterants in instant coffee premixes. The method development data covered chromatographic optimisation for better analyte separation and isomeric resolution, mass spectrometry optimisation for high sensitivity and sample preparation optimisation for high extraction recovery (RE) and low matrix effect (ME). The validation data covered specificity, linearity, range, accuracy, limit of detection, limit of quantification, precisions, ME, and RE. The optimisation and validation data presented here is related to the article: "Determination of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes using liquid chromatography-high-resolution mass spectrometry (LC-HRMS)" Mohd Yusop et al., 2019.

Matched MeSH terms: Limit of Detection

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