Displaying publications 1 - 20 of 37 in total

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  1. Abd Hamid MR, Tai ELM, Wan Hazabbah WH, Ramli N
    J Allergy Clin Immunol Pract, 2019 Apr;7(4):1308-1309.
    PMID: 30581132 DOI: 10.1016/j.jaip.2018.11.016
    Matched MeSH terms: Drug Hypersensitivity/diagnosis*
  2. Amstutz U, Shear NH, Rieder MJ, Hwang S, Fung V, Nakamura H, et al.
    Epilepsia, 2014 Apr;55(4):496-506.
    PMID: 24597466 DOI: 10.1111/epi.12564
    To systematically review evidence on genetic risk factors for carbamazepine (CBZ)-induced hypersensitivity reactions (HSRs) and provide practice recommendations addressing the key questions: (1) Should genetic testing for HLA-B*15:02 and HLA-A*31:01 be performed in patients with an indication for CBZ therapy to reduce the occurrence of CBZ-induced HSRs? (2) Are there subgroups of patients who may benefit more from genetic testing for HLA-B*15:02 or HLA-A*31:01 compared to others? (3) How should patients with an indication for CBZ therapy be managed based on their genetic test results?
    Matched MeSH terms: Drug Hypersensitivity/diagnosis
  3. Asha'ari ZA, Suhaimi Y, Yusof RA, Rushdan I, Maraina CH
    Med J Malaysia, 2011 Aug;66(3):202-6.
    PMID: 22111441 MyJurnal
    We compared a newer serum specific IgE (SSIgE) test with skin prick testing (SPT) in the diagnosis of allergy in Malaysia. Ninety newly diagnosed allergic patients were enrolled for both tests. Using SPT as a clinical gold standard, the sensitivity, specificity, positive, and negative predictive values (PPV, NPV) were calculated for SSIgE for each of the common allergens tested. The highest positive results for both SPT and SSIgE were for house dust mite and cat. Compared to SPT, SSIgE showed better sensitivity but poorer specificity, low PPV and good NPV in all the allergens tested. Significant positive correlation was seen between the diameter of wheal and flare of SPT and the SSIgE results.
    Matched MeSH terms: Hypersensitivity/diagnosis*
  4. Chaubal TV, Bapat RA, Patil PG, Shetty A
    Contact Derm., 2016 Oct;75(4):256-7.
    PMID: 27620128 DOI: 10.1111/cod.12625
    Matched MeSH terms: Latex Hypersensitivity/diagnosis
  5. Devaraj NK
    BMJ Case Rep, 2019 Feb 01;12(2).
    PMID: 30709894 DOI: 10.1136/bcr-2018-228355
    Matched MeSH terms: Shellfish Hypersensitivity/diagnosis*
  6. Gendeh BS, Murad S, Razi AM, Abdullah N, Mohamed AS, Kadir KA
    Otolaryngol Head Neck Surg, 2000 May;122(5):758-62.
    PMID: 10793361
    The aim of the study was to determine the incidence of food and house dust mite (HDM) allergy in patients with nasal congestion and rhinorrhea attending the Otorhinolaryngology Clinic, National University of Malaysia, Kuala Lumpur. This was a prospective matched, controlled study of patients skin prick tested with commercial food and common aeroallergens. The participants were 148 Malaysian adults with symptoms of nasal congestion and rhinorrhea and 113 adult Malaysian control subjects without rhinitis symptoms. The skin prick test (SPT) was used to evaluate 11 foods common to the Malaysian diet and 3 HDM inhalants. Forty-eight percent of the patients with rhinitis had positive SPT results to foods, compared with 4.4% of control subjects (P < 0.05). The most commonly implicated foods were shrimp (48%) and rice (30%), which are common in the Malaysian diet. Seventy-two percent of rhinitis patients had positive SPT results to HDM, compared with 22.2% of control subjects (P < 0.05). Patients with rhinitis also had significantly more gastrointestinal problems than control subjects (P < 0.05). The incidences of HDM and food allergy are significantly greater in Malaysian adults with rhinitis symptoms than in control subjects without rhinitis. The effect of avoidance or immunotherapy awaits further study.
    Study site: Otorhinolaryngology Clinic, Pusat Perubatan Universiti Kebangsaan Malaysia (PPUKM), Kuala Lumpur, Malaysia
    Matched MeSH terms: Food Hypersensitivity/diagnosis*
  7. Goh JC, Juliana J, Malina O, Ngah ZU, Norhafizalena O
    Trop Biomed, 2007 Jun;24(1):37-46.
    PMID: 17568376 MyJurnal
    Indoor fungal reservoirs, particularly airborne Penicillium species, were identified throughout the ventilation system of the building and dissemination of fungi from those reservoirs was found to be occurring all the time. The objectives of this study were to determine the association between air concentration of indoor mould (Penicillium) and allergy symptoms among office workers. The study design used in this research was a cross-sectional study. Risk factors were identified through the questionnaire survey. Office workers were selected based on the proximity of their workstations to the microbiological air sampler used for the mould sampling. Results from the current study suggests that individual susceptibility of exposed subjects might be influenced by several factors associated with mould exposure; for example, inhaled mycotoxins or volatile organic compounds, which may, in some complex way, affect the immune response. This study provides the much needed preliminary baseline data for developing guidelines with validated findings that will be of use for policy decisions in Malaysia regarding indoor air quality. Results from this study are recommended for use in planning and implementing control measures in order to reduce the exposure to indoor mould and promote healthy working environment among the workers.
    Matched MeSH terms: Hypersensitivity/diagnosis*
  8. Hajeb P, Selamat J
    Clin Rev Allergy Immunol, 2012 Jun;42(3):365-85.
    PMID: 22045217 DOI: 10.1007/s12016-011-8284-9
    Seafood is common item in the world diet; Asian countries have the highest rates of fish consumption in the world, which is higher than world average. Several studies have been conducted on the epidemiology and clinical characteristics of seafood allergy in different countries, and some of the fish and seafood allergens unique to those regions have been characterized. Review on published data showed that seafood allergy is very ubiquitous in some regions of the world. Fish and shellfish are the most common seafood that cause adverse allergic reactions among nations; the symptoms ranged from oral allergy syndromes to urticaria and anaphylaxis. The major identified allergens are parvalbumin in fish and tropomyosin in shellfish. Nevertheless, such studies are lacking from some regions with high fish and seafood consumption. Furthermore, the published data are mostly from small groups of populations, which large-scale epidemiological studies need to be performed.
    Matched MeSH terms: Food Hypersensitivity/diagnosis
  9. Hamilton RG, Adkinson NF
    J Allergy Clin Immunol, 1998 Sep;102(3):482-90.
    PMID: 9768592
    BACKGROUND: No characterized diagnostic natural rubber latex skin testing material is licensed for use in the United States.

    OBJECTIVE: We have conducted a multicenter clinical skin testing study to document the safety and diagnostic sensitivity and specificity of a candidate Hevea brasiliensis nonammoniated latex (NAL) extract. These data are intended to support the licensing of this reagent for the diagnosis of latex allergy in high-risk populations.

    METHODS: Three hundred twenty-four subjects (304 adults and 20 children) were classified by their clinical history as having latex allergy (LA group, 124 adults and 10 children) or having no latex allergy (NLA group, 180 adults and 10 children). All subjects provided blood samples and then received sequential puncture skin tests (PSTs) at 1, 100, or 1000 microg/mL protein with a bifurcated needle and NAL (Greer Laboratories) from Malaysian Hevea brasiliensis (clone 600) sap. A 2-stage glove provocation test was used to clarify latex allergy status of individuals with positive history/negative PST result and negative history/positive PST result mismatches.

    RESULTS: Twenty-four subjects (15%) originally designated as having LA on the basis of their initial clinical history were reclassified to the NLA group on the basis of a negative glove provocation test result. Of the 134 subjects with LA, 54 (40%) were highly sensitive to latex, with a positive PST result at 1 microg/mL NAL. The Greer NAL reagent produced a positive PST rate (sensitivity) of 95% and 99% in subjects with LA at 100 microg/mL and 1 mg/mL, respectively. The negative PST rate (specificity) in 190 subjects with a negative history with the NAL extract at 100 microg/mL and 1 mg/mL, was 100% and 96%, respectively. Immediately after the PST, mild systemic reactions (mainly pruritus) were recorded in 16.1 % of the adults in the LA group and 4.4% of the adults in the NLA group. No reactions required treatment with epinephrine. Only mild delayed reactions were observed in 9.6% (LA group) and 2.8% (NLA group) of subjects 24 to 48 hours after PST. Mean wheal and erythema diameters measured in the 10 children in the LA group with spina bifida at 100 microg/mL and 1 mg/mL were similar to those observed in the adults in the LA group, suggesting that children are not at increased risk for systemic reactions compared with adults.

    CONCLUSIONS: A suggestive clinical history is necessary but not sufficient for a definitive diagnosis of IgE-dependent latex allergy. These data support the safety and diagnostic efficacy of the Greer NAL, skin test reagent at 100 micro/mL and 1 mg/mL for confirmatory PSTs.

    Matched MeSH terms: Latex Hypersensitivity/diagnosis*
  10. Hamilton RG, Adkinson NF
    J Allergy Clin Immunol, 1996 Nov;98(5 Pt 1):872-83.
    PMID: 8939150
    BACKGROUND: Nonammoniated latex, ammoniated latex, and rubber glove extracts are the only sources of natural rubber (Hevea brasiliensis) latex that have potential for use as skin testing reagents in the diagnosis of latex allergy. Their diagnostic sensitivity and specificity as skin test reagents are unknown.

    OBJECTIVE: We conducted a phase 1/2 clinical study to examine the safety and diagnostic accuracy (sensitivity and specificity) of nonammoniated latex, ammoniated latex, and rubber glove extracts as skin test extracts to identify the most efficacious source material for future skin test reagent development.

    METHODS: Twenty-four adults not allergic to latex, 19 adults with hand dermatitis or pruritus, and 59 adults with a latex allergy were identified by clinical history. All provided blood and then received puncture skin tests and intradermal skin tests with nonammoniated latex, ammoniated latex, and rubber glove extracts from Malaysian H. brasiliensis latex by use of sequential titration. A glove provocation test and IgE anti-latex RAST were used to clarify positive history-negative skin test response and negative history-positive skin test response mismatches.

    RESULTS: All three extracts were biologically safe and sterile. After normalization to 1 mg/ml of total protein, all three extracts produced equivalent diagnostic sensitivity and specificity in puncture skin tests and intradermal skin tests at various extract concentrations. Optimal diagnostic accuracy was safely achieved at 100 micrograms/ml for intradermal skin tests (e.g., nonammoniated latex: puncture skin test sensitivity 96%, specificity 100%; intradermal skin test sensitivity 93%, specificity 96%). The presence of IgE antibody in skin was highly correlated with IgE anti-latex in serum (nonammoniated latex: r = 0.98, p < 0.001; ammoniated latex: r = 0.94, p < 0.001; rubber glove extract: r = 0.96, p < 0.001). All five available subjects with a positive history, negative skin test response, and absence of IgE antibody in serum had a negative glove provocation test response, indicating no clinical evidence of latex allergy. No systemic or large local allergic reactions were observed with puncture skin tests or intradermal skin tests.

    CONCLUSIONS: Equivalent diagnostic sensitivity and specificity were observed with the nonammoniated latex, ammoniated latex, and rubber glove extract skin test reagents after normalization for total protein; nonammoniated latex may be considered the reagent of choice on the basis of practical quality control and reproducibility considerations.

    Matched MeSH terms: Drug Hypersensitivity/diagnosis*
  11. Hamizan AW, Christensen JM, Ebenzer J, Oakley G, Tattersall J, Sacks R, et al.
    Int Forum Allergy Rhinol, 2017 01;7(1):37-42.
    PMID: 27530103 DOI: 10.1002/alr.21835
    BACKGROUND: Middle turbinate edema could be a characteristic feature of aeroallergen sensitization. In this study we sought to determine the diagnostic characteristics of middle turbinate edema as a marker of inhalant allergy.

    METHODS: A cross-sectional diagnostic study was performed on patients who had undergone nasal endoscopy and allergy testing. Allergy status was determined by positive serology or epicutaneous testing. Endoscopy was reviewed by blinded assessors for middle turbinate head edema. Appearance was graded as either normal, focal, multifocal, diffuse, or polypoid edema. Receiver-operator (ROC) analysis, likelihood ratio (LR), sensitivity, specificity, and positive predictive value (PPV) were determined.

    RESULTS: One hundred eighty-seven patients representing 304 nasal cavities were assessed (42% female, age 39.74 ± 14.7 years, 57% allergic). Diffuse edema (PPV 91.7%/LR = 8) and polypoid edema (PPV 88.9%/LR = 6.2) demonstrated the strongest association with inhalant allergy. Multifocal edema was used as a cut-off to represent inhalant allergy from ROC analysis, which demonstrated 94.7% specificity and 23.4% sensitivity. The PPV for multifocal was 85.1% and LR = 4.4.

    CONCLUSION: Middle turbinate edema is a useful nasal endoscopic feature to predict presence of inhalant allergy and, although not sensitive, has excellent PPV.

    Matched MeSH terms: Hypersensitivity/diagnosis*
  12. Ho TM, Murad S, Kesavapillai R, Singaram SP
    Asian Pac J Allergy Immunol, 1995 Jun;13(1):11-6.
    PMID: 7488338
    This study was conducted to determine the seasonal prevalence of allergies to house dust, D. pteronyssinus, D. farinae, cat fur, dog hair, mixed moulds, mixed grass pollens and American cockroach. A total of 314 patients with clinically suspected allergic rhinitis was examined by prick test using commercial preparations of the above allergens. Total serum IgE of the patients was determined by a Sandwich ELISA. Ninety-six percent of the patients tested positive to more than one allergen. Most were positive to a combination of 4 allergens. More than 70% of the patients were positive to house dust, D. pteronyssinus, D. farinae and cat fur. Analysis indicates that for an individual who tests positive for house dust, there is a very high risk of the person being allergic to the dust mites and cat fur too. Most of the allergens had 2 peak period of high positive PT rates; mixed moulds and mixed grass pollens had 3 peaks. There was significant positive correlation between the monthly positive PT rates against mixed moulds and mixed grass pollens with maximum daily mean temperature and mean temperature at 14.00 hours.
    Matched MeSH terms: Hypersensitivity/diagnosis
  13. Ho TM, DeBruynne J, Ahamad M, Darussamin H
    Asian Pac J Allergy Immunol, 1997 Sep;15(3):123-6.
    PMID: 9438543
    The MAST CLA system was evaluated against skin prick test (SPT) for diagnosis of allergies to house dust mites (Dermatophagoides pteronyssinus, Dermatophagoides farinae) and cats. Forty three asthmatic children were examined by SPT and MAST CLA. Chi-square analysis indicated significant association between SPT and MAST CLA results for the house dust mites but not for cats. The sensitivities of MAST CLA for house dust mites and cats were 100 and 25% respectively; specificities were all less than 50%. The efficiency of MAST CLA for detection of allergy to the house dust mites was 88% and 44% for cats. A significant linear correlation was found between SPT wheal size and MAST CLA grade for D. farinae but not for D. pteronyssinus and cats. It is concluded that the MAST CLA allergy system can be used to supplement SPT for diagnosis of allergies to house dust mites but not to cats.
    Matched MeSH terms: Hypersensitivity/diagnosis*
  14. Ho TM, Radha K, Shahnaz M, Singaram SP
    Med J Malaysia, 1993 Sep;48(3):308-12.
    PMID: 8183144
    An indirect enzyme linked immunosorbent assay (ELISA) was developed for the diagnosis of allergy to a house dust mite, Dermatophagoides farinae. The efficacy of the ELISA was then evaluated against a prick test using a commercial allergen. Eighty five suspected allergic rhinitis patients from the Otorhinolaryngology Department, Kuala Lumpur General Hospital, were tested with the ELISA and prick test. Prick test and ELISA results were positive in 84.7% and 80.0% of the patients respectively. The ELISA was found to have 87.5% sensitivity, 61.5% specificity, 92.6% positive predictive value, 47.1% negative predictive value, 7.4% false positive and 52.9% false negative. There was total agreement between the prick test and ELISA for prick test grades of 3+ and 4+. It is concluded that the ELISA is a useful assay for detection of individuals who are highly sensitive to D. farinae.
    Matched MeSH terms: Hypersensitivity/diagnosis*
  15. Ho TM, Yit YH, Husain M
    Asian Pac J Allergy Immunol, 1988 Dec;6(2):103-6.
    PMID: 3146256
    Allergy to Dermatophagoides pteronyssinus was determined in 61 rhinitis patients using prick test (PT), enzyme-immunoassay (EIA) and enzyme-linked immunosorbent assay (ELISA). A total of 43 patients tested positive with PT. Forty six patients were positive when tested with EIA and ELISA. With PT as standard test, EIA was found to have 83.7% sensitivity and 44.4% specificity; ELISA had 81.4% sensitivity and 38.9% specificity. There was a linear relationship between absorbance values obtained by EIA and ELISA. The performance time was 8 hours, 24 hours and 30 minutes for ELISA, EIA and PT respectively. The cost per test for ELISA, EIA and PT was US$ 0.20, US$ 5.20 and US$ 0.14 respectively. It was concluded that ELISA was more cost-effective than EIA should be used to supplement PT for a more complete diagnosis of allergy.
    Matched MeSH terms: Hypersensitivity/diagnosis
  16. Iyngkaran N, Yadav M, Boey CG
    Singapore Med J, 1995 Aug;36(4):393-6.
    PMID: 8919154
    Enterokinase has a critical role in initiating proteolytic digestion by hydrolysing the conversion of pancreatic trypsinogen into trypsin. The enzyme is synthesised by enterocytes of the proximal small intestine and initially incorporated into the brush border from where it is released into the intestinal lumen by the action of pancreatic secretions. The aim of the study was to analyse enterokinase activity in the duodenal mucosa of infants with diarrhoeal disease including cow's milk protein-sensitive enteropathy. Our observations show that the mean depletion of enterokinase was only 17% compared to 60-80% for other brush border enzymes like disaccharidases, peptidases and alkaline phosphatases in infants with diarrhoea. This suggests that enterokinase activity in the small bowel enteropathies may be dependent not only on the degree of mucosal damage specifically but also on the extent of damage to the goblet cell population where the enzyme is synthesised. Thus the enterokinase activity was reduced in acute and chronic diarrhoea with marked mucosal damage where significant reduction of goblet cell population was evident but the enzyme was relatively little affected when the mucosa was damaged mildly.
    Matched MeSH terms: Milk Hypersensitivity/diagnosis*
  17. Jambari NN, Liddell S, Martinez-Pomares L, Alcocer MJC
    PLoS One, 2021;16(4):e0249876.
    PMID: 33914740 DOI: 10.1371/journal.pone.0249876
    Ber e 1, a major Brazil nut allergen, has been successfully produced in the yeast Pichia pastoris expression system as homogenous recombinant Ber e 1 (rBer e 1) with similar physicochemical properties and identical immunoreactivity to its native counterpart, nBer e 1. However, O-linked glycans was detected on the P.pastoris-derived rBer e 1, which is not naturally present in nBer e 1, and may contribute to the allergic sensitisation. In this study, we addressed the glycosylation differences between P. pastoris-derived recombinant Ber e 1 and its native counterparts. We also determined whether this fungal glycosylation could affect the antigenicity and immunogenicity of the rBer e 1 by using dendritic cells (DC) as an immune cell model due to their role in modulating the immune response. We identified that the glycosylation occurs at Ser96, Ser101 and Ser110 on the large chain and Ser19 on the small polypeptide chain of rBer e 1 only. The glycosylation on rBer e 1 was shown to elicit varying degree of antigenicity by binding to different combination of human leukocyte antigens (HLA) at different frequencies compared to nBer e 1 when tested using human DC-T cell assay. However, both forms of Ber e 1 are weak immunogens based from their low response indexes (RI). Glycans present on rBer e 1 were shown to increase the efficiency of the protein recognition and internalization by murine bone marrow-derived dendritic cells (bmDC) via C-type lectin receptors, particularly the mannose receptor (MR), compared to the non-glycosylated nBer e 1 and SFA8, a weak allergenic 2S albumin protein from sunflower seed. Binding of glycosylated rBer e 1 to MR alone was found to not induce the production of IL-10 that modulates bmDC to polarise Th2 cell response by suppressing IL-12 production and DC maturation. Our findings suggest that the O-linked glycosylation by P. pastoris has a small but measurable effect on the in vitro antigenicity of the rBer e 1 compared to its non-glycosylated counterpart, nBer e 1, and thus may influence its applications in diagnostics and immunotherapy.
    Matched MeSH terms: Food Hypersensitivity/diagnosis
  18. Joo Chan C, Richardo T, Lim RLH
    Int Rev Immunol, 2018;37(6):279-290.
    PMID: 30638084 DOI: 10.1080/08830185.2018.1509967
    Peanut allergy is a hypersensitivity reaction with symptoms varying from mild to severe anaphylaxis, tends to be lifelong and very few are able to outgrow this allergy. The prevalence of peanut allergy is highest among the Western countries and over the past decade, a 3.5 fold increase in prevalence of peanut allergy was reported among children in the United States. Increasing prevalence has also been observed among the Asian countries. As with other food allergies, peanut allergy reduces quality of life for the affected individuals and the social and economy burden of healthcare for peanut allergy is substantial. To date, there is no effective treatment for peanut allergy and disease management is by avoidance or relieve of symptoms via administration of epinephrine. Peanut allergy is a type-1 hypersensitivity reaction due to specific IgE production by activated T-helper type 2 (TH2) cells. Studies on various immunotherapy routes such as oral immunotherapy (OIT), sublingual immunotherapy and epicutaneous immunotherapy trials using peanut have shown the ability to induce desensitisation, shifting the allergen-specific cytokine production away from a TH2 respond. In the recent years, lactic acid bacteria probiotics have been reported to down-regulate allergy due to its inherent immunomodulatory properties. Wild-type probiotic in combination with peanut proteins or recombinant probiotics harbouring peanut allergens have been explored for OIT due to its ability to down-regulate allergen-specific-IgE production and the TH2 responses, while increasing the beneficiary population of TH1 regulatory T cells (Treg). This review discusses the current strategies in immunotherapy for peanut allergy.
    Matched MeSH terms: Peanut Hypersensitivity/diagnosis
  19. Leecyous B, Bakhtiar F, Tang MM, Yadzir ZHM, Abdullah N
    Allergol Immunopathol (Madr), 2020 06 09;48(6):626-632.
    PMID: 32532468 DOI: 10.1016/j.aller.2020.01.006
    INTRODUCTION: Basophil activation test (BAT) and immunoassays are the most widely used in vitro tests to diagnose IgE-mediated allergic reactions to penicillin. However, studies to determine if one test is interdependent from another are limited.

    OBJECTIVE: The present study aimed to measure the agreement between BAT and immunoassay in diagnosis of penicillin allergy.

    METHOD: BAT was performed using penicillin G (Pen G), penicillin V (Pen V), penicilloyl-polylysine (PPL), minor determinant mix (MDM), amoxicillin (Amx) and ampicillin (Amp) in 25 patients. Immunoassay of total IgE (tIgE) and specific IgE (sIgE) antibodies to Pen G, Pen V, Amx and Amp were quantified. Skin prick test (SPT) using PPL-MDM, Amx, Amp and Clavulanic acid were also performed.

    RESULTS: Minimal agreement was observed between BAT and immunoassay (k=0.25). Of two BAT-positive patients, one patient is positive to Amx (59.27%, SI=59) and Amp (82.32%, SI=82) but sIgE-negative to all drug tested. This patient is also SPT-positive to both drugs. Another patient is BAT-positive to Pen G (10.18%, SI=40), Pen V (25.07%, SI=100) and Amp (19.52%, SI=79). In sIgE immunoassay, four patients were sIgE-positive to at least one of the drugs tested. The sIgE level of three patients was between low and moderate and they were BAT-negative. One BAT-positive patient had a high level of sIgE antibodies (3.50-17.5kU/L) along with relatively high specific to total IgE ratio ≥0.002 (0.004-0.007).

    CONCLUSIONS: The agreement between BAT and immunoassay is minimal. Performing both tests provides little increase in the sensitivity of allergy diagnosis work-up for immediate reactions to penicillin.

    Matched MeSH terms: Drug Hypersensitivity/diagnosis*
  20. Lew MH, Lim RL
    Appl Microbiol Biotechnol, 2016 Jan;100(2):661-71.
    PMID: 26411458 DOI: 10.1007/s00253-015-6953-y
    Current diagnostic tools for peanut allergy using crude peanut extract showed low predictive value and reduced specificity for detection of peanut allergen-specific immunoglobulin E (IgE). The Ara h 2.02, an isoform of the major peanut allergen Ara h 2, contains three IgE epitope recognition sequence of 'DPYSPS' and may be a better reagent for component resolve diagnosis. This research aimed to generate a codon-optimised Ara h 2.02 gene for heterologous expression in Escherichia coli and allergenicity study of this recombinant protein. The codon-optimised gene was generated by PCR using overlapping primers and cloned into the pET-28a (+) expression vector. Moderate expression of a 22.5 kDa 6xhistidine-tagged recombinant Ara h 2.02 protein (6xHis-rAra h 2.02) in BL21 (DE3) host cells was observed upon induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). The insoluble recombinant protein was purified under denaturing condition using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography and refolded by dialysis in decreasing urea concentration, amounting to a yield of 74 mg/l of expression culture. Matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) and immunoblot analysis confirmed the production of the recombinant 6xHis-rAra h 2.02. The refolded recombinant 6xHis-rAra h 2.02, with or without adjuvant, was able to elicit comparable level of allergen-specific IgE and IgG1 in sensitised Balb/c mice. In addition, the specific IgE antibodies raised against the recombinant protein were able to recognise the native Ara h 2 protein, demonstrating its allergenicity and potential as a reagent for diagnosis and therapeutic study.
    Matched MeSH terms: Peanut Hypersensitivity/diagnosis
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