Displaying publications 61 - 80 of 595 in total

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  1. Ben Hameid AS, Al-Sindi TA, Allow AK, Nafie EM, Alahmad BE, Faisal GG
    Oman Med J, 2019 May;34(3):238-243.
    PMID: 31110632 DOI: 10.5001/omj.2019.45
    Objectives: Estradiol (E2) plays an important role in the pathophysiology of ovarian hyperstimulation syndrome (OHSS). This study aimed to evaluate the effect of fenugreek seed aqueous (FSA) extract on serum E2 levels in a rat model of OHSS.

    Methods: A total of 34 female Sprague Dawley rats, aged 18 days old, weighing 40 to 45 g, were randomly divided into negative control, positive control, and treatment groups. A daily dose of 1500 mg/kg per body weight of FSA extract was administrated orally to rats in the treatment group for 13 days. On day eight of the study, OHSS was induced in both positive control and treated groups by subcutaneous injection of pregnant mare's serum gonadotropin 50 IU for four consecutive days, followed by human chorionic gonadotropin 25 IU on the fifth day. The effect of FSA extract was evaluated by measuring the concentration of serum E2 using the enzyme-linked immunosorbent assay.

    Results: FSA extract reduced serum E2 level significantly in the treated OHSS model (p-value < 0.050) compared to the positive control group.

    Conclusions: The finding has important implications on the development of female infertility adjuvant drugs for safe assisted reproduction technology cycles in terms of OHSS prevention.

    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  2. Bhartiya NM, Husain AA, Daginawala HF, Singh L, Kashyap RS
    Malays J Med Sci, 2020 Dec;27(6):15-26.
    PMID: 33447131 DOI: 10.21315/mjms2020.27.6.3
    Background: Human brucellosis is an important zoonotic disease of public health and often remains neglected owing to lack of sensitive and efficient diagnostic methods. This study evaluates diagnostic utility of in-house designed enzyme-linked immunosorbent assay (ELISA) using whole-cell antigens of Brucella abortus (B. abortus) S19 against the commercially available kits.

    Methods: A prospective cohort study involving different populations within the Vidarbha regions of Maharashtra, India was conducted through camps organised from May 2009 to October 2015. A total of 568 serum samples were collected from high-risk people recruited as study cohorts based on inclusion criteria, additional risk factors and clinical symptoms. Samples were evaluated by indirect ELISA using the whole-cell antigens of B. abortus. The results were compared with the commercially available IgG detection ELISA kit to ascertain the specificity and sensitivity of the developed test.

    Results: Fever, body ache, joint pain, lower back pain, loss of appetite and weight loss were major symptoms associated with the disease. With the cut-off of > 0.8, the positivity of brucellosis infection was at 12.32% (70/568) compared to 9.33% (53/568) as detected by the commercial kit. The in-house developed ELISA method yielded a sensitivity of 87.5% and specificity of 99.18% as compared to the commercial kits (sensitivity -80.30% and specificity -99.6%).

    Discussion: The B. abortus S19-derived whole-cell protein-based ELISA is rapid and cost-effective and can be used for screening brucellosis infection in lieu of the commercially available ELISA kits.

    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  3. Boo NY, Cheong KB, Cheong SK, Lye MS, Zulfiqar MA
    J Paediatr Child Health, 1997 Aug;33(4):329-34.
    PMID: 9323622
    OBJECTIVES: To compare the overall accuracy of the stable microbubble test (SM test) with measurement of level of surfactant protein A (SP-A) of tracheal aspirate for the diagnosis of respiratory distress syndrome (RDS).

    METHODOLOGY: Tracheal aspirates were obtained from neonates on ventilatory support. The SM test was carried out on specimens of tracheal aspirate immediately after collection. Levels of SP-A in tracheal aspirates were determined by enzyme-linked immunosorbent assay (ELISA) method. The results of the SM test and SP-A level of the tracheal aspirates were compared against the clinical diagnosis of RDS based on clinical, radiological and bacteriological findings.

    RESULTS: Both the median microbubble counts (6 microbubbles/mm2, range = 0-90) and median SP-A levels (100 micrograms/L, range = 0-67447) of infants with RDS were significantly lower than those of infants with no obvious lung pathology (P < 0.0001), and pneumonia (P < 0.0001). The SM test of tracheal aspirates had higher overall accuracy for the diagnosis of RDS than measurement of SP-A levels (94.6% vs 82.4%). When the receiver operating characteristic (ROC) curves of both tests for RDS were compared, the area under the ROC curve of the SM test was larger (0.9689) than that of the SP-A method (0.8965).

    CONCLUSIONS: This study showed that the SM test of tracheal aspirate was a useful bedside diagnostic test for RDS. It could be carried out at any time after birth on infants requiring ventilatory support.

    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  4. Cardosa MJ, Tio PH, Nimmannitya S, Nisalak A, Innis B
    PMID: 1298081
    The highly sensitive AFRIMS format IgM capture ELISA for the diagnosis of dengue virus infections requires the use of mouse brain derived hemagglutinins and consequently also the use of 20% acetone extracted normal human serum to eliminate high background. These reagents are not always easily available and we have thus compared the AFRIMS format with another published format which uses cell culture derived antigens (culture fluid, CF, format) in order to determine if it is reasonable to use cell culture derived antigens in situations where hemagglutinins and normal human serum are difficult to obtain. The study shows that using AFRIMS results as the reference point, the CF format described here has a sensitivity of 90% and a specificity of 96%.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  5. Cardosa MJ, Hah FL, Choo BH, Padmanathan S
    PMID: 8160055
    A dot enzyme immunoassay for determination of antibodies to Japanese encephalitis virus was designed for use as a field technique for the surveillance of Japanese encephalitis virus activity among domestic pigs. The test was compared with the neutralization test and the hemagglutination inhibition test and found to be more sensitive than the hemagglutination inhibition test and comparable to the neutralization test in sensitivity but more simple to perform than either the neutralization or the hemagglutination inhibition tests. An IgM capture ELISA for the determination of JEV specific porcine IgM was also utilized to determine current infection rates in pigs. The tests which do not involve the determination of specific IgM are better used for testing sentinel animals for providing clues as to the rate of transmission of JEV among pigs. IgM tests determining acute infection are less likely to be useful unless animals are tested very frequently or if a great number of animals are tested at any one time.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  6. Cardosa MJ, Choo BH, Zuraini I
    PMID: 1667957
    This study describes the status of viral encephalitis in Perak, Malaysia during the year 1990. In addition, 14 cases selected from Penang and Perak during the years 1989 and 1990 are presented, with data showing titers of neutralizing antibodies against Japanese encephalitis virus (JEV) and dengue 2 virus, titers of antibodies against JEV and dengue virus antigens as determined by DEIA, and a comparison of these with the presence of IgM to JEV and dengue virus. These data show that there probably is far more viral encephalitis due to JEV in Malaysia than the national figures reflect.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/standards
  7. Cardosa MJ, Zuraini I
    PMID: 1818383
    This study describes the use of an IgM capture ELISA using cell culture derived antigens and a polyclonal rabbit antiflavivirus antisera for the detection of dengue positive cases. The IgM capture ELISA is compared with the dot enzyme immunoassay and the results are discussed in the context of dengue endemicity.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/standards*
  8. Ch'ng ACW, Ahmad A, Konthur Z, Lim TS
    Methods Mol Biol, 2019;1904:377-400.
    PMID: 30539481 DOI: 10.1007/978-1-4939-8958-4_18
    Panning is a common process used for antibody selection from phage antibody libraries. There are several methods developed for a similar purpose, namely streptavidin mass spectrometry immunoassay (MSIA™) Disposable Automation Research Tips, magnetic beads, polystyrene immunotubes, and microtiter plate. The advantage of using a magnetic particle processor system is the ability to carry out phage display panning against multiple target antigens simultaneously in parallel. The system carries out the panning procedure using magnetic nanoparticles in microtiter plates. The entire incubation, wash, and elution process is then automated in this setup. The system also allows customization for the introduction of different panning stringencies. The nature of the biopanning process coupled with the limitation of the system means that minimal human intervention is required for the infection and phage packaging stage. However, the process still allows for rapid and reproducible antibody generation to be carried out.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  9. Ch'ng ACW, Konthur Z, Lim TS
    Methods Enzymol, 2020;630:159-178.
    PMID: 31931984 DOI: 10.1016/bs.mie.2019.10.023
    Directed evolution is a proven approach to fine tune or modify biomolecules for various applications ranging from research to industry. The process of evolution requires methods that are capable of not only generating genetic diversity but also to distinguish the variants of desired characteristics. One method that is synonymous with directed evolution of proteins is phage display. Here, we present a protocol describing the application of magnetic nanoparticles coupled with a processor to carry out the identification of monoclonal antibodies (mAbs) from a diverse antibody library via phage display. Target antigens are coupled to magnetic nanoparticles as the solid phase for the isolation of the binding mAbs via affinity. A gradual enrichment in clones would result in increasing ELISA readouts with increasing rounds of panning. During monoclonal level analysis, positivity can be deduced with comparison to background and controls. The biopanning process can also be adopted for the directed evolution of enzymes, scaffold proteins or even peptides.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods
  10. Ch'ng ACW, Hamidon NHB, Konthur Z, Lim TS
    Methods Mol Biol, 2018;1701:301-319.
    PMID: 29116512 DOI: 10.1007/978-1-4939-7447-4_16
    The application of recombinant human antibodies is growing rapidly mainly in the field of diagnostics and therapeutics. To identify antibodies against a specific antigen, panning selection is carried out using different display technologies. Phage display technology remains the preferred platform due to its robustness and efficiency in biopanning experiments. There are both manual and semi-automated panning selections using polystyrene plastic, magnetic beads, and nitrocellulose as the immobilizing solid surface. Magnetic nanoparticles allow for improved antigen binding due to their large surface area. The Kingfisher Flex magnetic particle processing system was originally designed to aid in RNA, DNA, and protein extraction using magnetic beads. However, the system can be programmed for antibody phage display panning. The automation allows for a reduction in human error and improves reproducibility in between selections with the preprogrammed movements. The system requires minimum human intervention to operate; however, human intervention is needed for post-panning steps like phage rescue. In addition, polyclonal and monoclonal ELISA can be performed using the semi-automated platform to evaluate the selected antibody clones. This chapter will summarize the suggested protocol from the panning stage till the monoclonal ELISA evaluation. Other than this, important notes on the possible optimization and troubleshooting are also included at the end of this chapter.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods
  11. Ch'ng WC, Saw WT, Yusoff K, Shafee N
    Acta Virol., 2011;55(3):227-33.
    PMID: 21978156
    Enterovirus 71 (EV71) is one of the viruses that cause hand, foot and mouth disease. Its viral capsid protein 1 (VP1), which contains many neutralization epitopes, is an ideal target for vaccine development. Recently, we reported the induction of a strong immune response in rabbits to a truncated VP1 fragment (Nt-VP1t) displayed on a recombinant Newcastle disease virus (NDV) capsid protein. Protective efficacy of this vaccine, however, can only be tested in mice, since all EV71 animal models thus far were developed in mouse systems. In this study, we evaluated the type of immune responses against the protein developed by adult BALB/c mice. Nt-VP1t protein induced high levels of VP1 IgG antibody production in mice. Purified VP1 antigen stimulated activation, proliferation and differentiation of splenocytes harvested from these mice. They also produced significant levels of IFN-γ, a Th1-related cytokine. Taken together, Nt-VP1t protein is a potent immunogen in adult mice and our findings provide the data needed for testing of its protective efficacy in mouse models of EV71 infections.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  12. Chaharaein B, Omar AR, Aini I, Yusoff K, Hassan SS
    Microbiol Res, 2009;164(2):174-9.
    PMID: 17336046
    Subtype-specific multiplex reverse transcription-polymerase chain reaction (RT-PCR) was developed to simultaneously detect three subtypes (H5, H7 and H9) of avian influenza virus (AIV) type A. The sensitivity of the multiplex RT-PCR was evaluated and compared to that of RT-PCR-enzyme-linked immunosorbent assay (ELISA) and conventional RT-PCR. While the sensitivity of the multiplex RT-PCR is as sensitive as the conventional RT-PCR, it is 10 times less sensitive than RT-PCR-ELISA. The multiplex RT-PCR is also as sensitive as the virus isolation method in detecting H9N2 from tracheal samples collected at day 3 and 5 post inoculation. Hence, the developed multiplex RT-PCR assay is a rapid, sensitive and specific assay for detecting of AIV subtypes.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  13. Chan M, Cheong TG, Kurunathan S, Chandrika M, Ledon T, Fando R, et al.
    Microb Pathog, 2010 Nov;49(5):211-6.
    PMID: 20558271 DOI: 10.1016/j.micpath.2010.06.001
    Cholera caused by the O139 serogroup still remains a public health concern in certain regions of the world and the existing O1 vaccines do not cross-protect cholera caused by this serogroup. An aminolevulinic acid (ALA) auxotroph vaccine candidate against the O139 serogroup, designated as VCUSM2, was recently developed. It was found to be immunogenic in animal model studies but showed mild reactogenic effects due to the presence of two intact copies of Vibrio cholerae toxin (CTX) genetic element. In the present study we have modified the ctx operon by systematic allelic replacement methodology to produce a mutant strain, designated as VCUSM14. This strain has two copies of chromosomally integrated and mutated ctxA gene, encoding immunogenic but not toxic cholera toxin A subunit (CT-A). The amino acids arginine and glutamic acid at position 7th and 112th, respectively, in CT-A of VCUSM14 were substituted with lysine (R7K) and glutamine (E112Q), respectively. Two copies of the ace and zot genes present in the ctx operon were also deleted. Cholera toxin-ELISA using GM1 ganglioside showed that the both wild type CT and mutated CT were recognized by anti-CT polyclonal antibodies. VCUSM14 produced comparatively less amount of antigenic cholera toxin when compared to the VCUSM2 and Bengal wild type strain. VCUSM14 did not elicit fluid accumulation when inoculated into rabbit ileal loops at doses of 10(6) and 10(8) CFU. The colonization efficiency of VCUSM14 was one log lower than the parent strain, VCUSM2, which can be attributed to the ALA auxotrophy and less invasive properties of VCUSM14. VCUSM14, thus a non-reactogenic auxotrophic vaccine candidate against infection by O139 V. cholerae.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  14. Chan SY, Kautner IM, Lam SK
    J Virol Methods, 1994 Oct;49(3):315-22.
    PMID: 7868649
    The potential of RT-PCR to rapidly diagnose dengue infections from both acute and convalescent phase patients' sera was evaluated. The RNA extraction method involved binding of the viral RNA to silica particles in the presence of high concentration of guanidine thiocyanate. The protocol that was established was sensitive enough to detect 40 plaque forming units per 100 microliter of serum and results could be obtained within one day. Results from this study indicate that clinical samples should be collected in the early acute phase of illness when anti-dengue antibodies were undetectable or of low titres to ensure a more reliable diagnosis.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  15. Chan WY, Selvarajah GT, Ajat M, Suzuki R, Tsukui T
    Vet Immunol Immunopathol, 2019 Jun;212:43-49.
    PMID: 31213251 DOI: 10.1016/j.vetimm.2019.05.002
    Canine atopic dermatitis (AD) is a chronic, inflammatory and pruritic allergic skin disease in dogs. House dust mites such as Dermatophagoides farinae are one of the known causative agents for the induction of canine AD worldwide. D. farinae protein Der f 2 is known as an important allergen involved in canine AD and recently, Zen-1 has also been identified as an allergenic protein. There is limited information on the prevalence and role of allergen sensitization to crude D. farinae extract (CDF), Der f 2 and Zen-1 among dogs diagnosed with AD in Malaysia. The aim of this study was to determine the proportion of CDF-, Der f 2- and Zen-1-specific reactive sera among dogs diagnosed with AD in Malaysia using an enzyme-linked immunosorbent assay (ELISA). Serum samples were collected from dogs diagnosed with AD from several veterinary clinics in Malaysia. The canine case records were retrieved and information on signalment, dermatological and non-dermatological histories, clinical presentation, food allergies, and exclusion of ectoparasitic, microbial and fungal skin infections were obtained through a survey form. All serum samples were evaluated to quantify the CDF-, Der f 2- and Zen-1-specific immunoglobulin E (IgE) levels. A total of 24.6%, 48.4% and 29.8% of dogs diagnosed with AD were positive for CDF-, Der f 2- and Zen-1-specific IgE, respectively. These results suggest that CDF-, Der f 2- and Zen-1 are important allergens that can contribute to AD in dogs in Malaysia, and serological testing can be performed to provide additional treatment options involving specific immunotherapies.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  16. Chandrasekaran S, Kennett L, Yeap PC, Muniandy N, Rani B, Mukkur TK
    Vet Microbiol, 1994 Aug 15;41(4):303-9.
    PMID: 7801530
    The relationship between the standard passive mouse protection test or serum antibody titres measured by indirect haemagglutination or enzyme-linked immunosorbent assays and active protection in buffaloes immunized with different types of haemorrhagic septicaemia bacterins was investigated. Groups of 2-3 buffaloes were immunized with the bacterins currently in use in Asia, viz., broth bacterin (BB), alum precipitated vaccine (APV) and oil adjuvant vaccine (OAV) either subcutaneously (BB, APV) or intramuscularly (OAV) and challenged subcutaneously with virulent organisms at different periods post-immunization. Although the passive mouse protection and indirect haemagglutination tests carried out with the pre-challenge sera from vaccinated buffaloes revealed no relationship with active protection in buffaloes, a relationship was observed between the ELISA antibody titres and protection. In contrast, a dose-response relationship was observed between the homologous active and passive mouse protection test.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/veterinary
  17. Chandrawathani P, Tsuji N, Kawazu S, Ishikawa M, Fujisaki K
    J Vet Med Sci, 1994 Oct;56(5):929-32.
    PMID: 7865596
    Cattle in Peninsular Malaysia were examined for evidence of infection with Babesia ovata, B. bigemina and B. bovis by an enzyme-linked immunosorbent assay for detection of antibody to the three Babesia species. All of the test samples when assayed with B. ovata antigen, resulted in low value indicating low probability of cattle infected with B. bigemina, 74.4% were positive for B. bovis and 72.6% were positive for both Babesia species. In addition, a serological survey with regard to age difference was carried out on a milk production farm. High reactivity antibody to B. bigemina and B. bovis was detected in calves less than 1 month of the age. The reactivity decreased in calves 1-3 months of the age. Then, the reactivity increased for both Babesia species in 6 months old calves. These results suggested that cattle infected with B. bigemina and B. bovis were widespread throughout Peninsular Malaysia and that both parasites might exist as an enzootical parasite.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  18. Chang MS, Matusop A, Sen FK
    PMID: 10774651
    Anopheles mosquitos were surveyed using three trapping technics in four longhouse settlements and their respectively farming zone in western Sarawak, Malaysia. The study area was mountainous with tropical rain forest. An. leucosphyrus and An. donaldi were predominant in the farm huts. An. tessellatus and An. subpictus were more abundant in the village settlements. In both ecotypes, human baited traps yielded a significantly greater proportion of Anopheles mosquito than CDC light traps and landing biting catches. Circumsporozoite antigen positively rate, mosquito survival rate and parasite rate showed that malaria transmission is more intense in farm huts than in longhouse settlements. The entomological inoculation rate of An. donaldi and An. leucosphyrus in farm huts was 0.035 and 0.023, respectively. No sporozoite infections were observed in the main settlements.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  19. Che-Amat A, Risalde MÁ, González-Barrio D, Ortíz JA, Gortázar C
    BMC Vet Res, 2016 Sep 05;12(1):184.
    PMID: 27596591 DOI: 10.1186/s12917-016-0825-2
    Diagnosing tuberculosis (TB) in farmed red deer (Cervus elaphus) is challenging and might require combining cellular and humoral diagnostic tests. Repeated skin-testing with mycobacterial purified protein derivatives (PPDs) might sensitize or desensitize the subjects to both kinds of diagnostic tools. We evaluated the effect of repeated (every 6 months) comparative tuberculin skin testing on skin test and ELISA responsiveness in farmed red deer hinds from a TB-free herd. Eighteen 8-month old hinds were inoculated with bovine and avian PPDs and the mitogen phytohaemagglutinin (PHA), as positive control and concurrently tested by ELISA for antibodies against avian (avian PPD, aPPD and protoplasmatic antigen 3, PPA3) and bovine antigens (bPPD and MPB70). Blood serum was also sampled three weeks after each skin testing round and tested for antibodies against aPPD and bPPD, in order to detect eventual antibody level boosts. Testing took place every six months from winter 2012 until winter 2015.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/veterinary
  20. Cheah TS, Sani RA, Chandrawathani P, Bahri S, Dahlan I
    Trop Anim Health Prod, 1999 Feb;31(1):25-31.
    PMID: 10399814
    An investigation into the epidemiology of Trypansoma evansi infection in crossbred dairy cattle was conducted for a period of 12 months on a dairy cattle farm in Penninsular Malaysia. The prevalence of parasitaemia was highest in lactating animals (13.4%), followed by those in the dry herd (8.8%), late pregnant animals (8.1%), early pregnant animals (4.7%), calves (0.3%) and heifers (0.2%). The prevalence of antigenaemia was highest in the lactating animals (54.7%), followed by that in dry animals (53.7%), heifers (51.1%), late pregnant animals (47.7%), early pregnant animals (46.5%) and calves (24.2%).
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/veterinary
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