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Applicability of Brugia malayi immune antibody library for the isolation of a human recombinant monoclonal antibody to Echinococcus granulosus antigen B

Rahumatullah A, Ahmad A, Noordin R, Lai JY, Baharudeen Z, Lim TS

Exp Parasitol, 2020 Dec;219:108029.
PMID: 33096112 DOI: 10.1016/j.exppara.2020.108029

Echinococcus granulosus is a worldwide zoonotic infection that causes human cystic echinococcosis (CE) or hydatid disease. The present study describes the isolation and production of a monoclonal antibody against recombinant AgB protein using the developed Human AntibodY Disease ENhanced (HAYDEN)-Filariasis library. The DNA sequences of the isolated clones were analyzed, followed by gene analysis and binding assays. Clone E1 showed a full-length sequence and represents the IgHV5-LV3 antibody gene family. The antibody protein yield was satisfactory, and it reacted specifically against rAgB. The novel E1 protein is potentially useful for the development of an antigen detection assay for CE. The ability of the Brugia malayi immune antibody library to isolate antibodies against Echinococcus granulosus antigens highlights the broad coverage of immune antibody libraries.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Application of PCR-ELISA in molecular diagnosis

Sue MJ, Yeap SK, Omar AR, Tan SW

Biomed Res Int, 2014;2014:653014.
PMID: 24971343 DOI: 10.1155/2014/653014

Polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) is an immunodetection method that can quantify PCR product directly after immobilization of biotinylated DNA on a microplate. This method, which detects nucleic acid instead of protein, is a much more sensitive method compared to conventional PCR method, with shorter analytical time and lower detection limit. Its high specificity and sensitivity, together with its semiquantitative ability, give it a huge potential to serve as a powerful detection tool in various industries such as medical, veterinary, and agricultural industries. With the recent advances in PCR-ELISA, it is envisaged that the assay is more widely recognized for its fast and sensitive detection limit which could improve overall diagnostic time and quality.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
Application of streptavidin mass spectrometric immunoassay tips for immunoaffinity based antibody phage display panning

Chin CF, Ler LW, Choong YS, Ong EB, Ismail A, Tye GJ, et al.

J Microbiol Methods, 2016 Jan;120:6-14.
PMID: 26581498 DOI: 10.1016/j.mimet.2015.11.007

Antibody phage display panning involves the enrichment of antibodies against specific targets by affinity. In recent years, several new methods for panning have been introduced to accommodate the growing application of antibody phage display. The present work is concerned with the application of streptavidin mass spectrometry immunoassay (MSIA™) Disposable Automation Research Tips (D.A.R.T's®) for antibody phage display. The system was initially designed to isolate antigens by affinity selection for mass spectrometry analysis. The streptavidin MSIA™ D.A.R.T's® system allows for easy attachment of biotinylated target antigens on the solid surface for presentation to the phage library. As proof-of-concept, a domain antibody library was passed through the tips attached with the Hemolysin E antigen. After binding and washing, the bound phages were eluted via standard acid dissociation and the phages were rescued for subsequent panning rounds. Polyclonal enrichment was observed for three rounds of panning with five monoclonal domain antibodies identified. The proposed method allows for a convenient, rapid and semi-automated alternative to conventional antibody panning strategies.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods
Application of the Serodia-HCV particle agglutination for the detection of antibodies to hepatitis C virus

Ooi BG, Sinniah M, Ismail S, Baharuddin R

Malays J Pathol, 1996 Dec;18(2):89-93.
PMID: 10879228

The Serodia-HCV Particle Agglutination (HCV-PA) for the detection of HCV antibodies was compared with the Enzyme Immunoassay Test (UBI HCV EIA) for possible in-house use. A total of 150 specimens were analysed using UBI HCV EIA and Serodia-HCV PA. Of these, 80 (53.3%) were both PA and EIA positive and 59 (39.3%) were negative by both techniques. Eleven sera (7.4%) were found to be EIA-positive but PA-negative. These 11 discordant sera were further tested by the LiaTek-HCV III Immunoassay (Organon Teknika). Ten were found to be line immunoassay negative and one was line immunoassay positive. Failure of the PA to detect the HCV positive serum meant that a small proportion of HCV antibody positives may be missed by the PA test. We conclude that (i) EIA should continue to be the first line screening test in our laboratory, (ii) PA with its 100% specificity could be a useful supplementary screen for all EIA-positive sera and finally (iii) line immunoassay could be used on sera to resolve discordant results in the EIA and PA assays.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Aptamer-17β-estradiol-antibody sandwich ELISA for determination of gynecological endocrine function

Huang Y, Zhang L, Li Z, Gopinath SCB, Chen Y, Xiao Y

Biotechnol Appl Biochem, 2021 Aug;68(4):881-888.
PMID: 33245588 DOI: 10.1002/bab.2008

17β-Estradiol-E2 (17β-E2) is a steroid hormone that plays a major role in the reproductive endocrine system and is involved in various processes, such as pregnancy, fertility, and menopause. In this study, the performance of an enzyme-linked immunosorbent assay (ELISA) for 17β-E2 quantification was enhanced by using a gold nanoparticle (GNP)-conjugated aptamer. An anti-17β-E2-aptamer-GNP antibody was immobilized on an amine-modified ELISA surface. Then, 17β-E2 was allowed to interact with and be sandwiched by antibodies. Aptamer-GNP conjugation was confirmed by colorimetric assays via the naked eye and UV-visible light spectroscopy. The detection limit based on a signal-to-noise ratio (S/N) of 3 was estimated to be 1.5 nM (400 pg/mL), and the linear range was 1.5-50 nM. Control experiments (without 17β-E2/with a complementary aptamer sequence/with a nonimmune antibody) confirmed the specific detection of 17β-E2. Moreover, 17β-E2 spiking of human serum did not interrupt the interaction between 17β-E2 and its antibody and aptamer. Thus, the developed ELISA can be used as an alternate assay for quantification of 17β-E2 and assessment of endocrine-related gynecological problems.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay

Toh SY, Citartan M, Gopinath SC, Tang TH

Biosens Bioelectron, 2015 Feb 15;64:392-403.
PMID: 25278480 DOI: 10.1016/j.bios.2014.09.026

The application of antibodies in enzyme-linked immunosorbent assay (ELISA) is the basis of this diagnostic technique which is designed to detect a potpourri of complex target molecules such as cell surface antigens, allergens, and food contaminants. However, development of the systematic evolution of Ligands by Exponential Enrichment (SELEX) method, which can generate a nucleic acid-based probe (aptamer) that possess numerous advantages compared to antibodies, offers the possibility of using aptamers as an alternative molecular recognition element in ELISA. Compared to antibodies, aptamers are smaller in size, can be easily modified, are cheaper to produce, and can be generated against a wide array of target molecules. The application of aptamers in ELISA gives rise to an ELISA-derived assay called enzyme-linked apta-sorbent assay (ELASA). As with the ELISA method, ELASA can be used in several different configurations, including direct, indirect, and sandwich assays. This review provides an overview of the strategies involved in aptamer-based ELASA.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay*
Aras Hormon Testosteron dan Progesteron dalam Kelawar Cynopterus brachyotis di Kampus Universiti Kebangsaan Malaysia (Concentration of Testosterone and Progesterone Homones in Cynopterus Brachyotis Bats in the Universiti Kebangsaan Malaysia Campus)

Kong KH, Zubaid A., Alias Bin Kamis

Sains Malaysiana, 1997;26.

Cynopterus brachyotis, a fruit, leaf and nectar eating bat is a common frugivorous bat in Southeast Asia. In the present study, changes in the reproductive hormones of these bats were measured. Bats were captured and blood samples were taken monthly for seven months (January to July 1997) in the surrounding areas of Universiti Kebangsaan Malaysia (UKM) campus in Bangi. The concentration of the reproductive hormones in the blood, such as testosterone (male) and progesterone (female) were measured using ELISA. The results showed that body weight and concentration of reproductive hormones was correlated with the monthly rainfall. Abundance of food during the rainy season was believed to be the important factor in influencing body weight gain and increase in reproductive hormone synthesis.
Cynopterus brachyotis merupakan kelawar frugivor pemakan buah-buahan, daun dan nektar yang lazim di Asia Tenggara. Matlamat penyelidikan ini ialah untuk menentukan aras hormon yang dirembeskan oleh gonad kelawar frugivor ini, Kelawar-kelawar ini telah ditangkap dan diambil sampel darah pada setiap bulan selama tujuh bulan (Januari hingga Julai 1997) di sekitar kampus Universiti Kebangsaan Malaysia (UKM) di Bangi. Aras hormon pembiakan, iaitu testosteron pada kelawar jantan dan progesteron pada betina ditentukan dengan menggunakan kaedah ELISA. Hasil kajian menunjukkan bahawa berat badan dan aras hormon pembiakan berkait rapat dengan jumlah taburan hujan bulanan. Kelimpahan sumber makanan pada masa taburan hujan yang tinggi dipercayai merangsang pertambahan berat badan dan sintesis hormon pembiakan pada kelawar ini.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Arterial stiffness, inflammatory and pro-atherogenic markers in gestational diabetes mellitus

Salmi AA, Zaki NM, Zakaria R, Nor Aliza AG, Rasool AH

VASA, 2012 Mar;41(2):96-104.
PMID: 22403127 DOI: 10.1024/0301-1526/a000171

This study aims to determine whether gestational diabetes mellitus (GDM) is associated with increased arterial stiffness, inflammatory and pro-atherogenic markers compared to age matched controls.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Association between Urinary Aflatoxin (AFM₁) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia

Sulaiman SH, Jamaluddin R, Sabran MR

Nutrients, 2018 Apr 07;10(4).
PMID: 29642443 DOI: 10.3390/nu10040460

Aflatoxin is a food contaminant and its exposure through the diet is frequent and ubiquitous. A long-term dietary aflatoxin exposure has been linked to the development of liver cancer in populations with high prevalence of aflatoxin contamination in foods. Therefore, this study was conducted to identify the association between urinary aflatoxin M₁ (AFM₁), a biomarker of aflatoxin exposure, with the dietary intake among adults in Hulu Langat district, Selangor, Malaysia. Certain food products have higher potential for aflatoxin contamination and these were listed in a Food Frequency Questionnaire, which was given to all study participants. This allowed us to record consumption rates for each food product listed. Concomitantly, urine samples were collected, from adults in selected areas in Hulu Langat district, for the measurement of AFM₁ levels using an ELISA kit. Of the 444 urine samples collected and tested, 199 were positive for AFM₁, with 37 of them exceeding the limit of detection (LOD) of 0.64 ng/mL. Cereal products showed the highest consumption level among all food groups, with an average intake of 512.54 g per day. Chi-square analysis showed that consumption of eggs (X² = 4.77, p = 0.03) and dairy products (X² = 19.36, p < 0.01) had significant associations with urinary AFM₁ but both food groups were having a phi and Cramer's V value that less than 0.3, which indicated that the association between these food groups' consumption and AFM₁ level in urine was weak.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Association between house dust mite (HDM) sensitisation and asthma control using skin prick test and HDM antigen specific IgE levels in saliva of Malaysian children

Teo, Keat Seong, Cheah, Cheong Wooi, Mak, Joon Wah

International e-Journal of Science, Medicine & Education, 2015;9(2):3-12.
MyJurnal

Background: Sensitisation to house dust mite (HDM) has been regarded as a major risk factor for development of asthma. This study was carried out to investigate the profiles of HDM sensitisation among Malaysian children with asthma.
Material and Methods: The association between HDM sensitisation and control and severity of asthma was investigated. The salivary HDM specific IgE levels were quantified in different grades of control and severity of asthma in 125 unselected asthmatic children aged 5-12 years old attending the asthma follow-up clinic in Hospital Tuanku Ja’afar Seremban. An additional 29 non-asthmatic patients were selected as control. The skin prick test to assess sensitisation to Dermatophagoides pteronyssinus (DP) and Dermatophagoides farinae (DF) was performed on all the participants. A questionnaire regarding the control and severity of asthmatic symptoms of the subject was administered. Saliva was collected by voluntary spitting and ELISA was used to quantify the IgE specific to HDM antigen.
Results: There was a significant association between sensitisation to DP and DF and the control of asthma. The association between DP sensitisation and severity of asthma just failed to reach a significant level although there is a clear trend for this. Significant association was found between DF sensitisation and severity. The HDM specific IgE in the saliva was significantly higher in asthmatic patients compared to non-asthmatic patients. There was no significant difference between the specific IgE levels in patients with different severity status of asthma.
Conclusion: Salivary IgE levels may not be an appropriate indicator of the patients’ asthmatic condition in this study. However, it can be concluded that there is significant association between the sensitisation of HDM and the control and severity of asthma.
Study site: Asthma clinic, Hospital Tuanku Jaafar, Seremban, Negeri Sembilan, Malaysia

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Association between respirable hexavalent chromium (Cr-VI) compounds with urinary β2-microglobulin level of welders in an automotive components manufacturing plant

Shamsul, B.S., Zakirah, M.

Journal of Occupational Safety and Health, 2014;11(1):23-32.
MyJurnal

The main objective of this study is to determine the association between respirable hexavalent chromium compounds with urinary β2-microglobulin levels among welders in an automotive components manufacturing plant. 49 welders and 39 workers involved in stamping process were selected as the exposed and the comparative group. β2-microglobulin is a protein renal tubular dysfunction marker that can indicate renal dysfunction caused by heavy metal. Air samples of worker’s breathing zone were collected using personal air sampling pump and filter papers. Filter papers were then diluted and analysed with Atomic Absorption Spectrophotometry (AAS). Workers’ urine samples were collected at the end of 8-hour work shift and analysed with β2-microglobulin ELISA Kit (IBL-Hamburg) and a microtiter reader. Meanwhile, creatinine levels were analysed with creatinine test strips and Reflotron®. A mean concentration of respirable hexavalent chromium compounds in air for the exposed group was 0.135 ± 0.043μg/m3 while for the non-exposed group was 0.124 ± 0.029μg/m3. The mean level of urinary β2-microglobulin per creatinine for the exposed group was 84.996 ± 39.246μg/g while that of the comparative group was 61.365 ± 21.609μg/g. The concentrations of respirable hexavalent chromium compounds were higher in the exposed group compared to the comparative group (Z=-2.444, p=0.015). β2-microglobulin level was also higher in the exposed group compared to the non-exposed group (t=3.821, p=

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Association of -174 G>C interleukin-6 gene polymorphism with interleukin-6 and c-reactive protein levels and obesity: A case-control study among people/residents of Western Indonesia

Pramudji H, Demes CM, Dewi K, Tasmini T, Ahmad HS

Med J Malaysia, 2019 Oct;74(5):400-404.
PMID: 31649216

BACKGROUND: Interleukin-6 (IL-6) and C-Reactive Protein (CRP) are mediators of inflammatory responses and increase in people who are obese . The increase of IL-6 and CRP levels is modified by polymorphism of -174 G>C IL-6 gene.
AIM: The purpose of this study was to investigate the relationship between -174 G>C IL-6 polymorphism gene on the level of IL-6 and CRP in the population of western Indonesia obese who are obese.
METHODS: In this study, we examined 178 subjects consisting of 89 who are obese with BMI> 25, and controls with BMI between 18.5 and 23. Fasting blood was taken from each subject for the examination of IL-6 and CRP levels by the ELISA method. Determination of genotype -174 G>C IL-6 gene was examined by Polymerase Chain reaction- Restriction Fragment Length Polymorphism (PCR-RFLP) methods.
RESULTS: The results of this study showed increased levels of IL-6 and CRP in the obese group compared to the controls. In the obese group, CC genotype had higher CRP and lower IL-6 levels than the GC and GG genotypes. The frequency of CC genotype in the obese group was 47.2% compared with 28.1% in controls and this genotype was considered a risk factor for obesity. Carriers of the C genotype as a dominant or a recessive model had greater risk of obesity.
CONCLUSION: It was concluded that the polymorphism - 174G>C IL-6 gene is a risk factor for obesity and is associated with increased levels of IL-6 and CRP in an obese group of the Western Indonesian ethnic population.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Association of annual hormonal profile with gonad maturity of mahseer (Tor tambroides) in captivity

Ismail MF, Siraj SS, Daud SK, Harmin SA

Gen Comp Endocrinol, 2011 Jan 1;170(1):125-30.
PMID: 20888822 DOI: 10.1016/j.ygcen.2010.09.021

Annual gonad hormonal profile of wild, matured mahseer (29 males and 23 female) averaging in weight between 0.95±0.26 and 1.19±0.23 kg for males and females, respectively, were investigated from November 2007 to November 2008 using enzyme-linked immunosorbent assay (ELISA) technique. Blood was collected from caudal vein, monthly and plasma separation by centrifugation was done to measure reproductive hormones: 17β-estradiol (E(2)), testosterone (T), and 11-keto-testosterone (11KT). Gonads were sampled for histology processing to observe their maturity. Highest T level in females and males was recorded at 0.22±0.016 and 0.88±0.014 ng/ml, respectively. The 11KT showed several peaks and the highest value was noted at 0.7±0.018 ng/ml in November 2008. The female E(2) initially was at 1.48±0.16 ng/ml and significantly increased (P<0.05) to 1.53±0.39 ng/ml in November 2008. Ovaries were laden with oocytes in several stages in all the samples while testes gonad showed a high level of spermatids throughout the year. Changes in plasma level of the gonadal hormones were correlated with the ovarian and testes maturities. In conclusion, the study suggests that mahseer can be categorized as asynchronized and multiple spawner. The information gathered is important for appropriate breeding and conservation programs of the Malaysian mahseer.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Association of anti-CLIC2 and anti-HMGB1 autoantibodies with higher disease activity in systemic lupus erythematosus patients

Syahidatulamali CS, Wan Syamimee WG, Azwany YN, Wong KK, Che Maraina CH

J Postgrad Med, 2017 9 2;63(4):257-261.
PMID: 28862243 DOI: 10.4103/jpgm.JPGM_499_16

BACKGROUND: Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by numerous autoantibodies. In this study, we investigated the presence of anti-chloride intracellular channel 2 (anti-CLIC2) and anti-high mobility group box 1 (anti-HMGB1) autoantibodies in SLE patients (n = 43) versus healthy controls ([HCs] n = 43), and their association with serological parameters (antinuclear antibody [ANA], anti-double-stranded DNA [anti-dsDNA], and C-reactive protein [CRP]) and disease activity using Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) score (active or inactive).
SETTINGS AND DESIGN: Case-control study at Rheumatology Clinic of Universiti Sains Malaysia Hospital.
SUBJECTS AND METHODS: The sera of SLE patients and HCs were tested for the presence of anti-CLIC2 and anti-HMGB1 autoantibodies using human recombinant proteins and ELISA methodologies. Other serological parameters were evaluated according to routine procedures, and patients' demographic and clinical data were obtained.
STATISTICAL ANALYSIS: Mann-Whitney U-test, Chi-square test, Fisher's exact test, and receiver operating characteristic analysis.
RESULTS: Anti-CLIC2 autoantibody levels were significantly higher in SLE patients compared to HCs (P = 0.0035), whereas anti-HMGB1 autoantibody levels were not significantly elevated (P = 0.7702). Anti-CLIC2 and anti-HMGB1 autoantibody levels were not associated with ANA pattern, anti-dsDNA, and CRP. Interestingly, SLEDAI score (≥6) was associated with anti-CLIC2 (P = 0.0046) and with anti-HMGB1 (P = 0.0091) autoantibody levels.
CONCLUSION: Our findings support the potential of using anti-CLIC2 autoantibodies as a novel biomarker for SLE patients. Both anti-CLIC2 and anti-HMGB1 autoantibody levels demonstrated potential in monitoring SLE disease activity.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods
Fulltext Attenuation of influenza virus infectivity with herbal-marine compound (HESA-A): an in vitro study in MDCK cells

Mehrbod P, Ideris A, Omar AR, Hair-Bejo M, Tan SW, Kheiri MT, et al.

Virol J, 2012;9:44.
PMID: 22340010 DOI: 10.1186/1743-422X-9-44

The influenza virus is still one of the most important respiratory risks affecting humans which require effective treatments. In this case, traditional medications are of interest. HESA-A is an active natural biological compound from herbal-marine origin. Previous studies have reported that the therapeutic properties of HESA-A are able to treat psoriasis vulgaris and cancers. However, no antiviral properties have been reported.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Autoantibodies to tetraspanins (CD9, CD81 and CD82) in demyelinating diseases

Miyaji K, Paul F, Shahrizaila N, Umapathi T, Yuki N

J Neuroimmunol, 2016 Feb 15;291:78-81.
PMID: 26857499 DOI: 10.1016/j.jneuroim.2015.12.012

Tetraspanin family proteins, CD9, CD81 and CD82 are expressed in the oligodendrocytes and Schwann cells. We investigated autoantibodies to tetraspanin proteins in patients with demyelinating diseases. Sera were collected from 119 multiple sclerosis patients, 19 neuromyelitis optica, 42 acute inflammatory demyelinating polyneuropathy, 23 chronic inflammatory demyelinating polyneuropathy and 13 acute motor axonal neuropathy as well as 55 healthy controls. Few multiple sclerosis and acute inflammatory demyelinating polyneuropathy patients had autoantibodies that were weakly reactive to CD9 or CD81 but the significance is unclear. It is unlikely that these autoantibodies are pathogenic or serve as potential biomarkers in demyelinating diseases.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Automatic diagnosis of tuberculosis disease based on Plasmonic ELISA and color-based image classification

AbuHassan KJ, Bakhori NM, Kusnin N, Azmi UZM, Tania MH, Evans BA, et al.

Annu Int Conf IEEE Eng Med Biol Soc, 2017 Jul;2017:4512-4515.
PMID: 29060900 DOI: 10.1109/EMBC.2017.8037859

Tuberculosis (TB) remains one of the most devastating infectious diseases and its treatment efficiency is majorly influenced by the stage at which infection with the TB bacterium is diagnosed. The available methods for TB diagnosis are either time consuming, costly or not efficient. This study employs a signal generation mechanism for biosensing, known as Plasmonic ELISA, and computational intelligence to facilitate automatic diagnosis of TB. Plasmonic ELISA enables the detection of a few molecules of analyte by the incorporation of smart nanomaterials for better sensitivity of the developed detection system. The computational system uses k-means clustering and thresholding for image segmentation. This paper presents the results of the classification performance of the Plasmonic ELISA imaging data by using various types of classifiers. The five-fold cross-validation results show high accuracy rate (>97%) in classifying TB images using the entire data set. Future work will focus on developing an intelligent mobile-enabled expert system to diagnose TB in real-time. The intelligent system will be clinically validated and tested in collaboration with healthcare providers in Malaysia.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Bacterial immunity and immunogenesis of normal human salivary IgA and serum IgG2 antiphospholipid autoantibody: a link?

Cheng HM, Sam CK

Immunol Lett, 1990 Oct;26(1):7-10.
PMID: 2276764

The anti-phospholipid antibody (aPL) in 26 heat-inactivated normal human sera (NHS) was tested for IgG subclass in ELISA. The specific antibody in NHS included all four IgG antibody subclasses, as well as IgA. The incidence of IgG subclasses ranged from 50% (13/26) for IgG1 to 92% (24/26) for IgG2. Specific IgA anti-phospholipid antibody (aPL) was detected by ELISA in 38% (28/73) of normal human saliva. The salivary IgA aPL bound preferentially to anionic phospholipids including cardiolipin, phosphatidylserine and phosphatidic acid but not to phosphatidylcholine or sphingomyelin. Unlike aPL in normal human sera, aPL in saliva was predominantly not associated with the previously described heat-labile inhibitor of aPL. This may indicate a role of salivary IgA aPL in local immunity by binding to cross-reactive bacterial cell surface components including phospholipids.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Biodynamics of hepatitis C virus in haemodialysis patients in Pahang

Mohammed Saad, A.M., Mohammed Imad, A.M., Aini, H.H., Seman, M.R.

International Medical Journal Malaysia, 2012;11(1):23-30.
MyJurnal

Introduction: HCV infection is frequent in patients undergoing maintenance haemodialysis, with prevalence between 8 and 10%. Hepatitis C has an adverse effect on both patient and graft survival in those who get renal transplants. There are relatively scarce reports on the natural fluctuation in viral load level in patients on chronic haemodialysis. Materials and Methods: This is a longitudinal short-term three months study, where 27 chronic haemodialysis patients infected with known HCV genotypes were recruited from seven haemodialysis centres in Pahang. Serum samples were collected monthly, both pre- and post-haemodialysis sessions, over a period of three months. Viral RNA was extracted from serum using QIAamp Viral RNA Extraction kit (Qiagen). The HCV viral load was measured using one-step reverse transcriptase qPCR (Applied Biosystems) targeting the 5`HCV non-coding region (5’UTR). The serum α-IFN level was measured using commercial ELISA kit (Amersham, UK). Six biochemical liver function tests (AST, ALP, TP, albumin, ALT and TB) were also done for all pre-haemodialysis samples. Results: All patients showed persistent low level viral load that varied significantly over the study period (p = 0.001). HCV genotype 1 viral load was significantly higher than that of genotype 3. Conclusion: No apparent correlation could be recognized between the viral loads and the corresponding interferon-alpha levels which were detectable in only a few patients during the period of study.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Biotin-Streptavidin Competition Mediates Sensitive Detection of Biomolecules in Enzyme Linked Immunosorbent Assay

Lakshmipriya T, Gopinath SC, Tang TH

PLoS One, 2016;11(3):e0151153.
PMID: 26954237 DOI: 10.1371/journal.pone.0151153

Enzyme Linked Immunosorbent Assay (ELISA) is the gold standard assay for detecting and identifying biomolecules using antibodies as the probe. Improving ELISA is crucial for detecting disease-causing agents and facilitating diagnosis at the early stages of disease. Biotinylated antibody and streptavidin-conjugated horse radish peroxide (streptavidin-HRP) often are used with ELISA to enhance the detection of various kinds of targets. In the present study, we used a competition-based strategy in which we pre-mixed free biotin with streptavidin-HRP to generate high-performance system, as free biotin occupies some of the biotin binding sites on streptavidin, thereby providing more chances for streptavidin-HRP to bind with biotinylated antibody. ESAT-6, which is a protein secreted early during tuberculosis infection, was used as the model target. We found that 8 fM of free biotin mixed with streptavidin-HRP anchored the higher detection level of ESAT-6 by four-fold compared with detection without free biotin (only streptavidin-HRP), and the limit of detection of the new method was 250 pM. These results suggest that biotin-streptavidin competition can be used to improve the diagnosis of analytes in other types of sensors.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Enzyme-Linked Immunosorbent Assay/standards

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