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Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk upon ultrasound-treatment

Ewe JA, Wan Abdullah WN, Bhat R, Karim AA, Liong MT

Ultrason Sonochem, 2012 Jan;19(1):160-73.
PMID: 21775184 DOI: 10.1016/j.ultsonch.2011.06.013

This study aimed at utilizing ultrasound treatment to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli (Lactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633, L. gasseri FTDC 8131) were treated with ultrasound (30 kHz, 100 W) at different amplitudes (20%, 60% and 100%) for 60, 120 and 180 s prior to inoculation and fermentation in biotin-soymilk. The treatment affected the fatty acids chain of the cellular membrane lipid bilayer, as shown by an increased lipid peroxidation (P<0.05). This led to increased membrane fluidity and subsequently, membrane permeability (P<0.05). The permeabilized cellular membranes had facilitated nutrient internalization and subsequent growth enhancement (P<0.05). Higher amplitudes and longer durations of the treatment promoted growth of lactobacilli in soymilk, with viable counts exceeding 9 log CFU/mL. The intracellular and extracellular β-glucosidase specific activities of lactobacilli were also enhanced (P<0.05) upon ultrasound treatment, leading to increased bioconversion of isoflavones in soymilk, particularly genistin and malonyl genistin to genistein. Results from this study show that ultrasound treatment on lactobacilli cells promotes (P<0.05) the β-glucosidase activity of cells for the benefit of enhanced (P<0.05) isoflavone glucosides bioconversion to bioactive aglycones in soymilk.

Matched MeSH terms: Biotin/metabolism*; Biotin/chemistry
Synthesis and Antitumor Evaluation of Glutathione Responsive Self-Immolative Disulphide Linked Camptothecin-Biotin Conjugate

Kaur A, Dhiman S, Lee HB, Sharma M

Anticancer Agents Med Chem, 2022;22(18):3182-3192.
PMID: 35469577 DOI: 10.2174/1871520622666220425114553

BACKGROUND: Camptothecin is a naturally occurring alkaloid obtained from the stem wood of the Chinese tree, Camptotheca acuminata. It exerts pharmacological effects due to its ability to selectively inhibit the type-I topoisomerase DNA nuclear enzyme. Several semisynthetic analogs of camptothecin have been synthesized to date possessing antitumor activity.
OBJECTIVE: Camptothecin (CPT) is one of the most promising anticancer drugs but it produces various side effects because of its non-selectivity towards cancer cells. To overcome these adverse effects, we synthesized biotin conjugate of camptothecin, which was linked via a self-immolative disulfide linker (CPT-SS-Biotin).
METHODS: Biotin conjugated camptothecin linked through a disulfide bond was synthesized following schemes, and the structural characterization was carried out. The stability and drug release studies were performed in the presence of glutathione (GSH) while in vitro studies were performed on 4T1 tumor cell lines. In vivo pharmacological investigation was done using an antitumor Wistar rat model.
RESULTS: The stability and drug release studies were performed in the presence of glutathione (GSH), and CPT-SSBiotin was found to be physiologically stable moiety and can only be cleaved in the presence of GSH to release free CPT. The CPT-SS-Biotin showed higher toxicity in the biotin-overexpressing 4T1 tumor cell line with a lower IC50 value (8.44 μM) compared to camptothecin alone (IC50 > 30 μM). CPT-SS-Biotin also showed 10.6% higher cellular uptake by cells in comparison to free camptothecin. The CPT-SS-Biotin was delivered to cells by binding to the biotin receptors on the cell surface, followed by energy-dependent endocytosis and internalization to cause cellular toxicity.
CONCLUSION: In-vivo tumor suppression studies and in vitro cell line studies along with serological parameters and histopathological studies showed that conjugate produced a high therapeutic effect and remarkably reduced toxic effects in comparison to free CPT. The results suggested that biotinylation of camptothecin via disulfide linker can be a safe and efficacious method in cancer therapeutics.

Matched MeSH terms: Biotin
Single-stranded DNA (ssDNA) production in DNA aptamer generation

Marimuthu C, Tang TH, Tominaga J, Tan SC, Gopinath SC

Analyst, 2012 Mar 21;137(6):1307-15.
PMID: 22314701 DOI: 10.1039/c2an15905h

The discovery that synthetic short chain nucleic acids are capable of selective binding to biological targets has made them to be widely used as molecular recognition elements. These nucleic acids, called aptamers, are comprised of two types, DNA and RNA aptamers, where the DNA aptamer is preferred over the latter due to its stability, making it widely used in a number of applications. However, the success of the DNA selection process through Systematic Evolution of Ligands by Exponential Enrichment (SELEX) experiments is very much dependent on its most critical step, which is the conversion of the dsDNA to ssDNA. There is a plethora of methods available in generating ssDNA from the corresponding dsDNA. These include asymmetric PCR, biotin-streptavidin separation, lambda exonuclease digestion and size separation on denaturing-urea PAGE. Herein, different methods of ssDNA generation following the PCR amplification step in SELEX are reviewed.

Matched MeSH terms: Biotin/metabolism; Biotin/chemistry
Fulltext Optic neuritis in a child with biotinidase deficiency: case report and literature review

Hayati AA, Wan-Hitam WH, Cheong MT, Yunus R, Shatriah I

Clin Ophthalmol, 2012;6:389-95.
PMID: 22457589 DOI: 10.2147/OPTH.S29048

Optic atrophy has often been reported in children with biotinidase deficiency. The visual prognosis is usually poor. This report is of a 6-year-old boy with an early onset of biotinidase deficiency who presented with acute profound visual loss in both eyes. Fundoscopy revealed swollen discs in both eyes, and the imaging was consistent with bilateral optic neuritis. He was treated with systemic corticosteroid, and commenced on oral biotin. The final visual outcome was promising.

Matched MeSH terms: Biotin; Biotinidase Deficiency
Effects of ultrasound on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus fermentum BT 8633 in biotin-supplemented soymilk

Ewe JA, Wan-Abdullah WN, Alias AK, Liong MT

Ultrason Sonochem, 2012 Jul;19(4):890-900.
PMID: 22305107 DOI: 10.1016/j.ultsonch.2012.01.003

This study aimed to evaluate the effects of ultrasound on Lactobacillus fermentum BT 8633 in parent and subsequent passages based on their growth and isoflavone bioconversion activities in biotin-supplemented soymilk. The treated cells were also assessed for impact of ultrasound on probiotic properties. The growth of ultrasonicated parent cells increased (P<0.05) by 3.23-9.14% compared to that of the control during fermentation in biotin-soymilk. This was also associated with enhanced intracellular and extracellular (8.4-17.0% and 16.7-49.2%, respectively; P<0.05) β-glucosidase specific activity, leading to increased bioconversion of isoflavones glucosides to aglycones during fermentation in biotin-soymilk compared to that of the control (P<0.05). Such traits may be credited to the reversible permeabilized membrane of ultrasonicated parent cells that have facilitated the transport of molecules across the membrane. The growing characteristics of first, second and third passage of treated cells in biotin-soymilk were similar (P>0.05) to that of the control, where their growth, enzyme and isoflavone bioconversion activities (P>0.05) were comparable. This may be attributed to the temporary permeabilization in the membrane of treated cells. Ultrasound affected probiotic properties of parent L. fermentum, by reducing tolerance ability towards acid (pH 2) and bile; lowering inhibitory activities against selected pathogens and reducing adhesion ability compared to that of the control (P<0.05). The first, second and third passage of treated cells did not exhibit such traits, with the exception of their bile tolerance ability which was inherited to the first passage (P<0.05). Our results suggested that ultrasound could be used to increase bioactivity of biotin-soymilk via fermentation by probiotic L. fermentum FTDC 8633 for the development of functional food.

Matched MeSH terms: Biotin/chemistry*
Bioconversion of isoflavones and the probiotic properties of the electroporated parent and subsequent three subcultures of Lactobacillus fermentum BT 8219 in biotin-soymilk

Ewe JA, Wan-Abdullah WN, Alias AK, Liong MT

J Microbiol Biotechnol, 2012 Jul;22(7):947-59.
PMID: 22580314

This study was aimed at an evaluation of the potential inheritance of electroporation effects on Lactobacillus fermentum BT 8219 through to three subsequent subcultures, based on their growth, isoflavone bioconversion activities, and probiotic properties, in biotin-supplemented soymilk. Electroporation was seen to cause cell death immediately after treatment, followed by higher growth than the control during fermentation in biotin-soymilk (P<0.05). This was associated with enhanced intracellular and extracellular beta-glucosidase specific activity, leading to increased bioconversion of isoflavone glucosides to aglycones (P<0.05). The growing characteristics, enzyme, and isoflavone bioconversion activities of the first, second, and third subcultures of treated cells in biotin-soymilk were similar to the control (P>0.05). Electroporation affected the probiotic properties of parent L. fermentum BT 8219, by reducing its tolerance towards acid (pH 2) and bile, lowering its inhibitory activities against selected pathogens, and reducing its ability for adhesion, when compared with the control (P<0.05). The first, second, and third subcultures of the treated cells showed comparable traits with that of the control (P>0.05), with the exception of their bile tolerance ability, which was inherited to the treated cells of the first and second subcultures (P<0.05). Our results suggest that electroporation could be used to increase the bioactivity of biotin-soymilk via fermentation with probiotic L. fermentum BT 8219, with a view towards the development of functional foods.

Matched MeSH terms: Biotin/metabolism
Improved sensitivity of detection by avidin-biotin complex (ABC) immunocytochemistry in Epstein-Barr virus serology

Thoe SY, Sam CK, Cheng HM, Prasad U

J Med Virol, 1989 Dec;29(4):311-4.
PMID: 2559955

Serum antibodies against Epstein-Barr virus (EBV)-determined antigens have traditionally been titrated by the indirect immunofluorescence (IIF) technique. The avidin-biotin complex (ABC) immunocytochemical technique was used to determine the serum levels of IgA against EBV viral capsid antigen (IgA/VCA) and IgA against EBV early antigen (IgA/EA) in sera of 106 nasopharyngeal carcinoma (NPC) patients prior to treatment and 100 normal individuals. The sensitivity of the ABC technique is enhanced by an amplification of the antigen-antibody reaction, which involves the binding of the enzyme-linked ABC to the second biotinylated antibody. There was a good correlation (r = 0.9988) between ABC and IIF-determined IgA/VCA-positive titres, with the ABC technique being more sensitive than IIF in the detection of IgA/VCA in NPC sera: 94% (99/106) and 76% (80/106), respectively. The frequency of IgA/EA reactivity in NPC sera was also markedly increased by immunodetection with the ABC technique as compared with IIF technique: 63% (69/106) and 28% (30/106) respectively. Both the immunocytochemical techniques were equally specific in discriminating between elevated serum titres of IgA/VCA and IgA/EA in NPC sera from normal human sera.

Matched MeSH terms: Biotin*
Exploring the additive bio-agent impacts upon ectoine production by Halomonas salina DSM5928T using corn steep liquor and soybean hydrolysate as nutrient supplement

Chen PW, Cui ZY, Ng HS, Chi-Wei Lan J

J Biosci Bioeng, 2020 Aug;130(2):195-199.
PMID: 32370929 DOI: 10.1016/j.jbiosc.2020.03.011

Ectoine production using inexpensive and renewable biomass resources has attracted great interest among the researchers due to the low yields of ectoine in current fermentation approaches that complicate the large-scale production of ectoine. In this study, ectoine was produced from corn steep liquor (CSL) and soybean hydrolysate (SH) in replacement to yeast extract as the nitrogen sources for the fermentation process. To enhance the bacterial growth and ectoine production, biotin was added to the Halomonas salina fermentation media. In addition, the effects addition of surfactants such as Tween 80 and saponin on the ectoine production were also investigated. Results showed that both the CSL and SH can be used as the nitrogen source substitutes in the fermentation media. Higher amount of ectoine (1781.9 mg L-1) was produced in shake flask culture with SH-containing media as compared to CSL-containing media. A total of 2537.0 mg L-1 of ectoine was produced at pH 7 when SH-containing media was applied in the 2 L batch fermentation. Moreover, highest amount of ectoine (1802.0 mg L-1) was recorded in the SH-containing shake flask culture with addition of 0.2 μm mL-1 biotin. This study demonstrated the efficacy of industrial waste as the nutrient supplement for the fermentation of ectoine production.

Matched MeSH terms: Biotin/metabolism
Fulltext Screen Printed Carbon Electrode Based Electrochemical Immunosensor for the Detection of Dengue NS1 Antigen

Parkash O, Yean CY, Shueb RH

Diagnostics (Basel), 2014;4(4):165-80.
PMID: 26852684 DOI: 10.3390/diagnostics4040165

An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes (SPCEs) for the detection of the dengue NS1 antigen was developed in this study. Monoclonal anti-NS1 capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. Subsequently, a direct sandwich enzyme linked immunosorbent assay (ELISA) format was developed and optimized. An anti-NS1 detection antibody conjugated with horseradish peroxidase enzyme (HRP) and 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H₂O₂) was used as an enzyme mediator. Electrochemical detection was conducted using the chronoamperometric technique, and electrochemical responses were generated at -200 mV reduction potential. The calibration curve of the immunosensor showed a linear response between 0.5 µg/mL and 2 µg/mL and a detection limit of 0.03 µg/mL. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. In conclusion, this immunosensor is a promising technology for the rapid and convenient detection of acute dengue infection in real serum samples.

Matched MeSH terms: Biotin
Fulltext Halal analysis of raw materials, ingredients and finished bakery products using PCR and gene chip southern-hybridization for detection of porcine DNA

Norrakiah, A.S., Shahrul Azim, M. G., Sahilah, A.M., Abdul Salam, B.

International Food Research Journal, 2015;22(5):1883-1887.
MyJurnal

This study was conducted to investigate the sensitivity and detection of porcine DNA in raw materials, ingredients and finished bakery products by polymerase chain reaction (PCR) - southern hybridization on chip analysis. A total of 20 samples (n=20* 3) with three replicates for each samples were obtained from a bakery factory located in Bangi, Selangor from January to December 2012. The sensitivity level of PCR-southern hybridization on chip was 0.001 ng. The species-specific oligonucleotide primers used in PCR-southern hybridization were targeted on the mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene sequence, namely cty b biotin-labeled oligonucleotide primers. The amplicon from PCR amplification was 276 bp in size. None of the raw materials, ingredients and finished bakery product samples was positive towards porcine DNA, except for the positive control. The results in the present study demonstrated that the PCR- southern-hybridization technique on the gene chip (OliproTM Porcine gene chip) is a sensitive tool for monitoring the porcine component in highly processed ingredients and finished bakery products.

Matched MeSH terms: Biotin
Endothelial replicative senescence delayed by the inhibition of MTORC1 signaling involves MicroRNA-107

Khor ES, Wong PF

Int J Biochem Cell Biol, 2018 Aug;101:64-73.
PMID: 29857052 DOI: 10.1016/j.biocel.2018.05.016

Accumulation of senescent endothelial cells can contribute to endothelium dysfunction. Suppression of MTOR signaling has been shown to delay senescence but the mechanism that underpins this effect, particularly one that involves miRNAs, remains to be further defined. This study sought to identify miRNAs involved in MTORC1-mediated inhibition of replicative senescence in endothelial cells. Pre-senescent HUVECs were prolonged treated with low dose rapamycin (1 nM), an MTOR inhibitor. Rapamycin treatment down-regulated the phosphorylated MTOR, RPS6 and 4EBP1 expressions, which confirmed MTORC1 suppression. Prolonged low dose rapamycin treatment has significantly reduced the percentage of senescence-associated beta galactosidase (SA-β gal) positively stained senescent cells and P16INK4A expression in these cells. On the contrary, the percentage of BrdU-labelled proliferating cells has significantly increased. RPTOR, a positive regulator of MTORC1 was knockdown using RPTOR siRNA to inhibit MTORC1 activation. RPTOR knockdown was evidenced by significant suppressions of RPTOR mRNA and protein expression levels. In these cells, the expression of miR-107 was down-regulated whereas miR-145-5p and miR-217 were up-regulated. Target gene prediction revealed PTEN as the target of miR-107 and this was confirmed by biotin pull-down assay. Over-expression of miR-107 has decreased PTEN expression, increased MTORC1 activity, induced cell cycle arrest at G0/G1 phase and up-regulated P16INK4A expression but mitigated tube formation. Collectively, our findings revealed that delayed endothelial replicative senescence caused by the inhibition of MTORC1 activation could be modulated by miR-107 via its influence on PTEN.

Matched MeSH terms: Biotin
Biotin-thiamine responsive basal ganglia disease: a retrospective review of the clinical, radiological and molecular findings of cases in Kuwait with novel variants

Aburezq M, Alahmad A, Alsafi R, Al-Tawari A, Ramadan D, Shafik M, et al.

Orphanet J Rare Dis, 2023 Sep 05;18(1):271.
PMID: 37670342 DOI: 10.1186/s13023-023-02888-y

BACKGROUND: Biotin-thiamine-responsive basal ganglia disease (BTBGD) is a rare autosomal recessive neurometabolic disorder that is caused by biallelic pathogenic SLC19A3 variants and is characterized by subacute encephalopathy associated with confusion, convulsions, dysphagia, dysarthria, or other neurological manifestations.
METHODS: A retrospective review of the data registry in Kuwait Medical Genetics Center for all cases diagnosed clinically and radiographically and confirmed genetically with BTBGD.
RESULTS: Twenty one cases from 13 different families were diagnosed with BTBGD in Kuwait. Most cases (86%) presented with confusion, dystonia, convulsions, or dysarthria, while three individuals were diagnosed pre-symptomatically during familial targeted genetic screening. Symptoms resolved completely within 2-week of treatment in two-thirds of the symptomatic cases but progressed in six of them to a variety of severe symptoms including severe cogwheel rigidity, dystonia and quadriparesis due to delayed presentation and management. Neuroradiological findings of the symptomatic cases revealed bilateral central changes in the basal ganglia. Two novel homozygous missense SLC19A3 variants were detected in a Kuwaiti and a Jordanian individuals, in addition to the previously reported Saudi founder homozygous variant, c.1264A > G; p.(Thr422Ala) in the remaining cases. Age of diagnosis ranged from newborn to 32 years, with a median age of 2-3 years. All cases are still alive receiving high doses of biotin and thiamine.
CONCLUSION: This is the first study reporting the phenotypic and genotypic spectrum of 21 individuals with BTBGD in Kuwait and describing two novel SLC19A3 variants. BTBGD is a treatable neurometabolic disease that requires early recognition and treatment initiation. This study highlights the importance of performing targeted molecular testing of the founder variant in patients presenting with acute encephalopathy in the region.

Matched MeSH terms: Biotin
Rapid growth and osteogenic differentiation of mesenchymal stem cells isolated from human bone marrow

Shima WN, Ali AM, Subramani T, Mohamed Alitheen NB, Hamid M, Samsudin AR, et al.

Exp Ther Med, 2015 Jun;9(6):2202-2206.
PMID: 26136960

Mesenchymal stem cells (MSCs) are involved in bone formation in the embryo, bone repair and remodeling. The differentiation of these cells is a complex multistep pathway that involves discrete cellular transitions and is similar to that which occurs during hematopoiesis. MSCs have self-renewal capacity without differentiation in long-term culture. In the present study, MSCs were isolated from human bone marrow and characterized by the presence of cluster of differentiation 105 marker using the labeled streptavidin biotin method. The MSCs were cultured in Dulbecco's modified Eagle's medium supplemented with fetal bovine serum, ascorbic acid, β-glycerol phosphate and dexamethasone to differentiate into osteoblasts. Biological in vitro analysis showed the rapid proliferation of the MSCs. Further evaluation of specific osteogenic markers using von Kossa staining and the alkaline phosphate assay demonstrated that the MSCs were stimulated to differentiate into osteoblast-lineage cells. This mesengenic potential indicated that the bone marrow-derived cells were multipotent MSCs. The findings of this study show that bone marrow can be a legitimate source of MSCs for the production of osteoblasts for utilization in bone replacement therapy.

Matched MeSH terms: Biotin
Fulltext Pre-eclampsia: is it all in the placenta?

Singh HJ

Malays J Med Sci, 2009 Jan;16(1):7-15.
PMID: 22589643 MyJurnal

Hypertensive disorders of pregnancy complicate almost 7 - 10 % of all pregnancies. The dyad of hypertension and proteinuria after 20 weeks of gestation is referred to as pre-eclampsia. It is a major cause of maternal morbidity and mortality and is also associated with increased perinatal problems. Despite intensive research over the years the exact cause of pre-eclampsia remains unknown. Nevertheless, information gleaned from published studies point to the placenta as the probable pathogenetic focus of pre-eclampsia, as the disease usually resolves within 24 - 48 hours after delivery of the placenta. Although the precise involvement of the placenta in pre-eclampsia remains unclear there are indications that the trophoblastic invasion of the uterine spiral arteries is abnormal in women who develop pre-eclampsia. This impaired invasion leads to decreased placental perfusion and ultimately to placental hypoxia. The distressed or ischaemic placenta then secretes a factor(s) into the maternal circulation, which cause/s widespread endothelial cell dysfunction characterized by vasospasm, activation of coagulation system and organ ischaemia. The cause of the defective cytotrophoblastic invasion of the spiral arteries and the link between placental ischaemia and generalized maternal endothelial dysfunction remain unknown. Although the placenta appears to have a major role in the pathogenesis of pre-eclampsia, evidence also suggests that factors like maternal genetic predisposition, dietary, environmental and behaviour, which surface during the stress of pregnancy might also be involved in the development of pre-eclampsia. It is known that not all women with poor cytotrophoblast invasion develop pre-eclampsia and not all women with preeclampsia show poor cytotrophoblast invasion. Over the years, a number of potential risk factors associated with the development of pre-eclampsia are being recognized and it might be appropriate now to develop some preventative strategies based upon the available information.

Matched MeSH terms: Biotin
Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk by electroporation

Ewe JA, Wan-Abdullah WN, Alias AK, Liong MT

Int J Food Sci Nutr, 2012 Aug;63(5):580-96.
PMID: 22149599 DOI: 10.3109/09637486.2011.641940

This study aimed at utilizing electroporation to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli were treated with different pulsed electric field strength (2.5, 5.0 and 7.5 kV/cm) for 3, 3.5 and 4 ms prior to inoculation and fermentation in biotin-soymilk at 37°C for 24 h. Electroporation triggered structural changes within the cellular membrane of lactobacilli that caused lipid peroxidation (p 9 log CFU/ml after fermentation in biotin-soymilk (p biotin-soymilk (p biotin-soymilk with increased bioactive aglycones.

Matched MeSH terms: Biotin/metabolism*
Human papilloma virus DNA-biomarker analysis for cervical cancer: Signal enhancement by gold nanoparticle-coupled tetravalent streptavidin-biotin strategy

Lv Q, Wang Y, Su C, Lakshmipriya T, Gopinath SCB, Pandian K, et al.

Int J Biol Macromol, 2019 Aug 01;134:354-360.
PMID: 31078598 DOI: 10.1016/j.ijbiomac.2019.05.044

Human papillomavirus (HPV) is a double-standard DNA virus, as well as the source of infection to the mucous membrane. It is a sexually transmitted disease that brings the changes in the cervix cells. Oncogenes, E6 and E7 play a pivotal role in the HPV infection. Identifying these genes to detect HPV strains, especially a prevalent HPV16 strain, will bring a great impact. Among different sensing strategies for pathogens, the dielectric electrochemical biosensor shows the potential due to its higher sensitivity. In this research, HPV16-E7 DNA sequence was detected on the carbodiimidazole-modified interdigitated electrode (IDE) surface with the detection limit of 1 fM. To enhance the sensitivity, the target sequence was conjugated on gold nanoparticle (GNP) and attained detection to the level of 10 aM. This produced ~100 folds improvement in detecting HPV16-E7 gene and 4 folds increment in the current flow. The stability of HPV16-E7 DNA sequences on GNP was verified by the salt-induced GNP aggregation. The current system has shown the higher specificity by comparing against non-complementary and triple-mismatched DNA sequences of HPV16-E7. This demonstration in detecting HPV16-E7 using dielectric IDE sensing system with a higher sensitivity can be recommended for detecting a wide range of disease-causing DNA-markers.

Matched MeSH terms: Biotin/chemistry*
Enzyme-linked amperometric electrochemical genosensor assay for the detection of PCR amplicons on a streptavidin-treated screen-printed carbon electrode

Yean CY, Kamarudin B, Ozkan DA, Yin LS, Lalitha P, Ismail A, et al.

Anal Chem, 2008 Apr 15;80(8):2774-9.
PMID: 18311943 DOI: 10.1021/ac702333x

A general purpose enzyme-based amperometric electrochemical genosensor assay was developed wherein polymerase chain reaction (PCR) amplicons labeled with both biotin and fluorescein were detected with peroxidase-conjugated antifluorescein antibody on a screen-printed carbon electrode (SPCE). As a proof of principle, the response selectivity of the genosensor was evaluated using PCR amplicons derived from lolB gene of Vibrio cholerae. Factors affecting immobilization, hybridization, and nonspecific binding were optimized to maximize sensitivity and reduce assay time. On the basis of the background amperometry signals obtained from nonspecific organisms and positive signals obtained from known V. cholerae, a threshold point of 4.20 microA signal was determined as positive. Under the optimum conditions, the limit of detection (LOD) of the assay was 10 CFU/mL of V. cholerae. The overall precision of this assay was good, with the coefficient of variation (CV) being 3.7% using SPCE and intermittent pulse amperometry (IPA) as an electrochemical technique. The assay is sensitive, safe, and cost-effective when compared to conventional agarose gel electrophoresis, real-time PCR, and other enzyme-linked assays for the detection of PCR amplicons. Furthermore, the use of a hand-held portable reader makes it suitable for use in the field.

Matched MeSH terms: Biotin/chemistry
Detection of influenza A and B neutralizing antibodies in vaccinated ferrets and macaques using specific biotin-streptavidin conjugated antibodies

Sirskyj D, Weltzin R, Golshani A, Anderson D, Bozic J, Diaz-Mitoma F, et al.

J Virol Methods, 2010 Feb;163(2):459-64.
PMID: 19913054 DOI: 10.1016/j.jviromet.2009.11.014

Several critical factors of an influenza microneutralization assay, utilizing a rapid biotin-streptavidin conjugated system for detecting influenza virus subtypes A and B, are addressed within this manuscript. Factors such as incubation times, amount of virus, cell seeding, sonication, and TPCK trypsin were evaluated for their ability to affect influenza virus neutralization in a microplate-based neutralization assay using Madin-Darby canine kidney (MDCK) cells. It is apparent that the amount of virus used in the assay is the most critical factor to be optimized in an influenza microneutralization assay. Results indicate that 100xTCID(50) of influenza A/Solomon Islands/03/2006 (H1N1) virus overloads the assay and results in no, to low, neutralization, in both ferret and macaque sera, respectively, whereas using 6xTCID(50) resulted in significantly improved neutralization. Conversely, strong neutralization was observed against 100xTCID(50) of B/Malaysia/2506/04 virus. In this manuscript the critical factors described above were optimized and the results indicate that the described biotin-streptavidin conjugated influenza microneutralization assay is a rapid and robust method for detecting the presence of functional, influenza virus-neutralizing antibodies.

Matched MeSH terms: Biotin/pharmacology
Fulltext The use of an in-house biotin-avidin linked immunosorbent assay to detect Aspergillus antigens in sera of immunocompromised patients

Abdul Samad S, Yusoff H, Fadilah SAW

Med J Malaysia, 2001 Mar;56(1):32-8.
PMID: 11503293

A biotin-avidin-linked immunosorbent assay was developed to detect Aspergillus antigens in sera of immunocompromised patients. The assay was based on a double antibody sandwich ELISA using polyclonal antibodies raised against water-soluble antigens of Aspergillus fumigatus. Aspergillus antigens were positive in sera of 9 of 16 (56%) patients who were studied prospectively and in 13 of 73 (19%) patients studied retrospectively. The 9 prospectively studied patients who were antigen positive were febrile neutropenic hematological malignancy patients who exhibited a high risk of acquiring invasive aspergillosis.

Matched MeSH terms: Biotin
Use of avidin-biotin-glucose oxidase complex to detect antimalarial antibody in serum by light microscopy

Lee M, Lambros C

Am J Trop Med Hyg, 1988 Aug;39(2):145-9.
PMID: 3044152

An immunohistochemical assay was developed combining an avidin-biotin-glucose oxidase complex procedure (ABC-GO) with light microscopy to detect specific antibody against Plasmodium falciparum. Thin blood films were prepared from culture material of P. falciparum and fixed with acetone. Antibody was detected by successive incubations with test serum, biotinylated goat antihuman antibody, avidin-biotin-glucose oxidase complex, and glucose oxidase substrate. In the presence of reactive serum, a blue precipitate formed on the parasites and could be visually observed with a 40x objective. Sera from patients with single infections for P. vivax or P. ovale were unreactive. No cross-reactivity was observed with sera from patients with rheumatoid arthritis, filariasis, amebiasis, schistosomiasis, dengue, scrub typhus, leptospirosis, or toxoplasmosis. The sensitivity of ABC-GO is comparable to that of the indirect fluorescent antibody test.

Matched MeSH terms: Biotin

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