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  1. A Brugia malayi antigen specifically recognized by infected individuals
    Rahmah N, Anuar AK, A'shikin AN, Lim BH, Mehdi R, Abdullah B, et al.
    Biochem Biophys Res Commun, 1998 Sep 29;250(3):586-8.
    PMID: 9784388
    Western blot analyses were performed on 444 serum specimens: 40 sera from microfilaraemic individuals, 10 sera from elephantiasis patients, 24 treated individuals, 50 sera from residents of endemic areas without anti-filarial IgG4 antibodies (endemic normals), 20 sera from amicrofilaraemic individuals with high anti-filarial IgG4 antibodies, 200 sera from healthy city-dwellers (non-endemic samples), and 100 sera from soil-transmitted helminth-infected individuals. Phast electrophoresis system was used to electrophorese Brugia malayi soluble adult worm antigen on 10-15% SDS-PAGE gradient gels followed by electrophoretic transfer onto PVDF membranes. Membrane strips were then successively incubated with blocking solution, human sera, and monoclonal anti-human IgG4 antibody-HRP, with adequate washings done in between each incubation step. Luminol chemiluminescence detection was then used to develop the blots. An antigenic band with the MW of approximately 37 kDa was found to be consistently present in the Western blots of all microfilaraemic sera, all amicrofilaraemic sera with high titres of anti-filarial IgG4 antibodies, some treated patients, and some elephantiasis patients. The antigen did not occur in immunoblots of individuals with other helminthic infections, normal endemic individuals, and city dwellers. Therefore the B. malayi antigen of with the MW of approximately 37 kDa demonstrated specific reactions with sera of B. malayi-infected individuals and thus may be useful for diagnostic application.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  2. Generation and selection of naïve Fab library for parasitic antigen: Anti-BmSXP antibodies for lymphatic filariasis
    Omar N, Hamidon NH, Yunus MH, Noordin R, Choong YS, Lim TS
    Biotechnol Appl Biochem, 2018 May;65(3):346-354.
    PMID: 28833498 DOI: 10.1002/bab.1591
    Phage display has been applied successfully as a tool for the generation of monoclonal antibodies (mAbs). Naive antibody libraries are unique as they are able to overcome several limitations associated with conventional mAb generation methods like the hybridoma technology. Here, we performed an in vitro selection and generation of Fab antibodies against Brugia malayi SXP protein (BmSXP), a recombinant antigen for the detection of lymphatic filariasis. We developed a naïve multi ethnic Fab antibody library with an estimated diversity of 2.99 × 109 . The antibody library was used to screen for mAbs against BmSXP recombinant antigen. Soluble monoclonal Fab antibodies against BmSXP were successfully isolated from the naïve library. The Fab antibodies obtained were expressed and analyzed to show its binding capability. The diversity obtained from a pool of donors from various ethnic groups allowed for a diverse antibody library to be generated. The mAbs obtained were also functional in soluble form, which makes it useful for further downstream applications. We believe that the Fab mAbs are valuable for further studies and could also contribute to improvements in the diagnosis of filariasis.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  3. Purification of recombinant antigen BmSXP used in panLF rapid kit for lymphatic filariasis
    Khoo TK, Noordin R, Santhanam A
    Indian J Exp Biol, 2012 Apr;50(4):256-64.
    PMID: 22611913
    A rapid antibody detection test is very useful for the detection of lymphatic filariasis, especially for certification and surveillance of post-mass drug administration. panLF Rapid kit is suitable for this purpose since it can detect all species of lymphatic filaria. It is based on the detection of anti-filarial IgG4 antibodies that react with recombinant B. malayi antigens, BmR1 and BmSXP. There is an increase demand for the test due to its attributes of being rapid, sensitive and specific results, as well as its field-applicability. The main aim of this paper is to obtain high recovery and purity of recombinant antigen BmSXP via a modified method of immobilized metal affinity chromatography (IMAC). The highest product yield of 11.82 mg/g dry cell weight (DCW) was obtained when IMAC was performed using the optimized protocol of 10 mM imidazole concentration in lysis buffer, 30 mM imidazole concentration in wash buffer, and 10 column volume wash buffer containing 300 mM salt concentration. This gave a 54% protein recovery improvement over the manufacturer's protocol which recorded a product yield of only 7.68 mg/g DCW. The recovered BmSXP recombinant antigen showed good western blot reactivity, high sensitivity (31/32, 97%) and specificity (32/32, 100%) in ELISA, thus attesting to its good purity and quality.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  4. Fulltext Antibodies to somatic L3 antigen not protective against Brugia malayi infection
    Noordin R, Abdullah KA, Azahri NA, Ramachandran CP
    Southeast Asian J Trop Med Public Health, 1999 Dec;30(4):678-81.
    PMID: 10928359
    Western blot analysis of infective larvae (L3) antigen of Brugia malayi were performed on 200 sera from six groups of individuals: 36 samples from B. malayi microfilaremic individuals; 10 samples from individuals with elephantiasis; 50 and 20 samples from amicrofilaremic individuals in a B. malayi endemic area with no anti-filarial IgG4 antibodies (towards microfilaria and adult worm antigens) and samples with high titres of the anti-filarial IgG4 antibodies respectively; 50 samples from non-endemic normals and 34 samples from geohelminth-infected individuals. After protein transfer, PVDF membrane strips were successively incubated with blocking solution, human sera, monoclonal anti-human IgG4 antibody-HRP and developed with luminol chemiluminescence substrate. 28/36 (78%), 1/10 (10%) and 16/20(80%) of sera from individuals with microfilariae, elephantiasis and amicrofilaremic individuals with high titers of anti-filarial IgG4 antibodies respectively recognized L3 antigenic epitopes; the dominant and consistent antigenic bands were of approximately MW 43 kDa, 14 kDa, 15 kDa and 59 kDa. The rest of the sera were unreactive. This study showed that microfilaremics may or may not mount a notable antibody response to somatic L3 antigens, thus lending evidence that antibody response to this antigen is not protective against establishment of Brugia malayi infection.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  5. Changes in complement C3 levels following treatment of patients with Brugia malayi infection
    Yadav M
    Southeast Asian J Trop Med Public Health, 1989 Jun;20(2):183-7.
    PMID: 2609207
    Serum IgG levels and complement C3 levels were assayed on Day 0, 1, 3-4, 7 and 56-70 post-treatment with diethylcarbamizine citrate (DEC) in a series to 26 patients with Brugia malayi infection and 6 volunteers without infection. On treatment, the microfilariae were cleared from the blood within 24 hours. The eosinophils decreased dramatically on Day 1 post-treatment but increased rapidly by Day 4 to 7 and then dropped to normal levels in 45 days. The serum IgG mean levels decreased briefly following treatment with DEC but then returned to original levels. However, the complement C3 levels gradually increased over the 2 months period of study reaching statistical significance levels (p less than 0.01) in patients with initial high blood microfilariae. The observation suggests that Brugia malayi infection probably induces a high rate of synthesis of complement C3 and this process continued in the post-treatment phase. Since, DEC treatment did not cause a decrease in complement C3 with the elimination of blood microfilariae, it would appear that the complement C3 is consumed following antibody attachment to the microfilariae as they enter the blood circulation.
    Matched MeSH terms: Elephantiasis, Filarial/immunology
  6. Fulltext Laboratory detection of strongyloidiasis: IgG-, IgG4 - and IgE-ELISAs and cross-reactivity with lymphatic filariasis
    Norsyahida A, Riazi M, Sadjjadi SM, Muhammad Hafiznur Y, Low HC, Zeehaida M, et al.
    Parasite Immunol., 2013 May-Jun;35(5-6):174-9.
    PMID: 23448095 DOI: 10.1111/pim.12029
    Enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of IgG, IgG4 and IgE antibodies against Strongyloides stercoralis. A commercial ELISA (IVD Research, USA) was also used, and the sensitivities and specificities of the four assays were determined. Serum samples from 26 patients with S. stercoralis infection and 55 patients with other infections or no infection were analysed. Sensitivities of the IgG4 , IgG, IgE and IgG (IVD) assays were 76.9%, 84.6%, 7.7% and 84.6%, respectively, while the specificities were 92.7%, 81.8%, 100% and 83.6%, respectively. If filariasis samples were excluded, the specificities of the IgG4 -ELISA and both IgG-ELISAs increased to 100% and 98%, respectively. A significant positive correlation was observed between IgG- and IgG4 -ELISAs (r = 0.4828; P = 0.0125). IgG- and IgG- (IVD) ELISAs (r = 0.309) were positively correlated, but was not significant (P = 0.124). Meanwhile there was no correlation between IgG4 - and IgG- (IVD) ELISAs (r = 0.0042; P = 0.8294). Sera from brugian filariasis patients showed weak, positive correlation between the titres of antifilarial IgG4 and the optical densities of anti-Strongyloides IgG4 -ELISA (r = 0.4544, P = 0.0294). In conclusion, the detection of both anti-Strongyloides IgG4 and IgG antibodies could improve the serodiagnosis of human strongyloidiasis. Furthermore, patients from lymphatic filariasis endemic areas who are serologically diagnosed with strongyloidiasis should also be tested for filariasis.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  7. Potential use of IgG2-ELISA in the diagnosis of chronic elephantiasis and IgG4-ELISA in the follow-up of microfilaraemic patients infected with Brugia malayi
    Rahmah N, Anuar AK, Karim R, Mehdi R, Sinniah B, Omar AW
    Biochem Biophys Res Commun, 1994 Nov 30;205(1):202-7.
    PMID: 7999024
    Sera from fifty subjects with different presentations of Brugian filariasis and from common soil-transmitted helminth infections were tested for specific anti-filarial IgG and its subclasses. Anti-filarial IgG, IgG1 and IgG3 showed cross-reactivities with soil-transmitted helminthic infections and no significant differences in optical densities among the various groups of filarial patients. In comparison with other groups of subjects, IgG4-ELISA of sera from microfilaraemic patients and some previously microfilaraemic patients showed a significant increase in optical density readings, while IgG2-ELISA showed elevated optical density readings in sera of patients with chronic elephantiasis. Therefore IgG2-ELISA is potentially useful in the diagnosis of brugian chronic elephantiasis while IgG4-ELISA may be beneficial for follow-up diagnosis of treated microfilaraemic patients.
    Matched MeSH terms: Elephantiasis, Filarial/immunology
  8. Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Rahumatullah A, Lim TS, Yunus MH, Noordin R
    Am J Trop Med Hyg, 2019 08;101(2):436-440.
    PMID: 31162018 DOI: 10.4269/ajtmh.19-0034
    Lymphatic filariasis is a mosquito-borne parasitic disease responsible for morbidity and disability that affects 1.2 billion people worldwide, mainly the poor communities. Currently, filarial antigen testing is the method of choice for the detection of bancroftian filariasis, and to date, there are two commonly used tests. In the present study, a recently reported recombinant monoclonal antibody (5B) specific to BmSXP filarial antigen was used in developing an ELISA for the detection of circulating filarial antigen in sera of patients with bancroftian filariasis. The performance of the ELISA was evaluated using 124 serum samples. The ELISA was positive with all sera from microfilaremic bancroftian filariasis patients (n = 34). It also showed 100% diagnostic specificity when tested with sera from 50 healthy individuals and 40 patients with other parasitic diseases. The developed assay using the novel 5B recombinant monoclonal antibody could potentially be a promising alternative antigen detection test for bancroftian filariasis.
    Matched MeSH terms: Elephantiasis, Filarial/immunology
  9. Fulltext Laboratory Evaluation of a Rapid IgG4 Antibody Test (BLF Rapid™) for Bancroftian Filariasis
    Noordin R, Yunus MH, Robinson K, Won KY, Babu S, Fischer PU, et al.
    Am J Trop Med Hyg, 2018 12;99(6):1587-1590.
    PMID: 30350768 DOI: 10.4269/ajtmh.18-0566
    At the end phase of the Global Programme to Eliminate Lymphatic Filariasis, antibody testing may have a role in decision-making for bancroftian filariasis-endemic areas. This study evaluated the diagnostic performance of BLF Rapid™, a prototype immunochromatographic IgG4-based test using BmSXP recombinant protein, for detection of bancroftian filariasis. The test was evaluated using 258 serum samples, comprising 96 samples tested at Universiti Sains Malaysia (in-house) and 162 samples tested independently at three international laboratories in the USA and India, and two laboratories in Malaysia. The independent testing involved 99 samples from Wuchereria bancrofti microfilaria or antigen positive individuals and 63 samples from people who were healthy or had other infections. The in-house evaluation showed 100% diagnostic sensitivity and specificity. The independent evaluations showed a diagnostic sensitivity of 84-100% and 100% specificity (excluding non-lymphatic filarial infections). BLF Rapid has potential as a surveillance diagnostic tool to make "Transmission Assessment Survey"-stopping decisions and conduct post-elimination surveillance.
    Matched MeSH terms: Elephantiasis, Filarial/immunology
  10. Seroprevalence of lymphatic filariasis among migrant workers in Peninsular Malaysia
    Noordin R, Mohd Zain SN, Yunus MH, Sahimin N
    Trans R Soc Trop Med Hyg, 2017 08 01;111(8):370-372.
    PMID: 29206992 DOI: 10.1093/trstmh/trx062
    Background: Malaysia aims to eliminate lymphatic filariasis (LF) by the year 2020, thus the potential threat of LF from migrant workers needs to be investigated.

    Methods: Brugian and bancroftian filariasis among 484 migrant workers from six countries were investigated using rapid tests based on detection of specific IgG4 antibodies against BmR1 (Brugia Rapid) and BmSXP recombinant antigens.

    Results: The seroprevalence of brugian filariasis was very low; however, bancroftian filariasis was notable among workers from India, Nepal and Myanmar.

    Conclusion: Malaysia is not endemic for Wuchereria bancrofti, but harbors the vectors for the parasite, thus the results showed that migrant workers should be monitored for this infection.

    Matched MeSH terms: Elephantiasis, Filarial/immunology
  11. Advances in immunology and immunopathology of lymphatic filariasis
    Mak JW
    Southeast Asian J Trop Med Public Health, 1993;24 Suppl 2:76-81.
    PMID: 7973952
    The lymphatic filarial parasites which affect about 90 million people worldwide have similar host-parasite relationships in man. They are all able to survive, reproduce and cause chronic infections if they can successfully evade the protective responses of the host. Studies to investigate the wide spectrum of clinical manifestations of the infection even among those living in similar endemic areas and with presumed equal exposure to infective larvae, have been hampered by the lack of animal models showing similar host-parasite responses. The recent use of the nude mouse infected with Brugia spp, and the leaf-monkey (Presbytis spp) infected with B. malayi or Wuchereria spp for the study of immune responses and the associated pathology of these infections, has elucidated some of the host protective immune responses as well as the associated immunopathological reactions. The successfully entrenched parasite elicits minimal reactions and pathology, but with the onset of effective host responses, whether assisted by chemotherapy, development of protective immunity or both, severe inflammatory responses may occur. The role of such immune mediated response in determining subsequent pathology will probably be dependent on the frequency and duration of these episodes, but these have yet to be defined. Prenatal and perinatal sensitization by filarial antigens are postulated to result in tolerance and/or modification of immune responses to subsequent infections. A role for genetic predisposition to certain clinical outcomes, for example, the development of elephantiasis, has been postulated but needs further study. Advances have also been achieved in defining those parasite antigens/products involved in eliciting or suppressing protective and other immune responses.(ABSTRACT TRUNCATED AT 250 WORDS)
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  12. Delineation of BmSXP antibody V-gene usage from a lymphatic filariasis based immune scFv antibody library
    Rahumatullah A, Ahmad A, Noordin R, Lim TS
    Mol Immunol, 2015 Oct;67(2 Pt B):512-23.
    PMID: 26277276 DOI: 10.1016/j.molimm.2015.07.040
    Phage display technology is an important tool for antibody generation or selection. This study describes the development of a scFv library and the subsequent analysis of identified monoclonal antibodies against BmSXP, a recombinant antigen for lymphatic filariasis. The immune library was generated from blood of lymphatic filariasis infected individuals. A TA based intermediary cloning approach was used to increase cloning efficiency for the library construction process. A diverse immune scFv library of 10(8) was generated. Six unique monoclonal antibodies were identified from the 50 isolated clones against BmSXP. Analysis of the clones showed a bias for the IgHV3 and Vκ1 (45.5%) and IgHV2 and Vκ3 (27.3%) gene family. The most favored J segment for light chain is IgKJ1 (45.5%). The most favored D and J segment for heavy chain are IgHD6-13 (75%) and IgHJ3 (47.7%). The information may suggest a predisposition of certain V genes in antibody responses against lymphatic filariasis.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  13. Fulltext Duration of detection of anti-BmR1 IgG4 antibodies after mass-drug administration (MDA) in Sarawak, Malaysia
    Noordin R, Muhi J, Md Idris Z, Arifin N, Kiyu A
    Trop Biomed, 2012 Mar;29(1):191-6.
    PMID: 22543621 MyJurnal
    The detection rates of brugian filariasis in three regions of Sarawak namely Central, North and South after three courses of mass drug administration (MDA) from year 2004 to 2006 was investigated. A recombinant BmR1 antigen-based IgG4 detection test, named Brugia Rapid and night blood smear for microfilaria (mf) detection were used. All three regions recorded a sharp fall in mf positive rates after a year post-MDA. Meanwhile Brugia Rapid positive rates declined more gradually to 3.8% and 5.6% of the pre-MDA levels in the Central and North regions, respectively. This study showed that in filariasis endemic areas in Sarawak, anti-filarial IgG4 antibodies to BmR1, as detected by the Brugia Rapid test, were positive for one to two years after mf disappearance.
    Matched MeSH terms: Elephantiasis, Filarial/immunology*
  14. Detection of circulating antigens and parasite specific antibodies in filariasis
    Abdullah WO, Oothuman P, Yunus H
    Southeast Asian J Trop Med Public Health, 1993;24 Suppl 2:31-6.
    PMID: 7973943
    In Peninsular Malaysia, only Wuchereria bancrofti and Brugia malayi are reported to cause human filariasis. Brugia pahangi infects many of the same animal hosts as the zoonotically transmitted subperiodic B. malayi. There is a well-recognized need for improved diagnostic techniques for lymphatic filariasis. Parasite antigen detection is a promising new approach, and it will probably prove to be more sensitive and specific than clinical, microscopic and antibody-based serological methods. We recently generated monoclonal antibodies (MAb XC3) from in vitro culture products of adult B. pahangi (B.p. IVP). Filarial antigenemia was quantitated in various hosts including the sera from 6 Malaysian Aborigines with acute lymphatic filariasis. In hosts infected with brugian filariasis and dirofilariasis, antigenemia was scored ranging from 90 ng/ml to 960 ng/ml. None of the control animal and human sera had antigenemia above 90 ng/ml. In addition, MAb XC3 and B.p. IVP were applied in several seroepidemiological surveys among household cats in Kuala Selangor in order to correlate information gathered for future studies of possible cases of human infection. Out of the 81 cats surveyed, 10 (12.35%) and 5 (6.17%) were parasitologically positive for B. pahangi and B. malayi, respectively. However, 21 (25.92%) were antigenemia positive when serologically investigated with MAb XC3. Antifilarial antibodies to B.p. IVP by direct ELISA showed very high cross-reactivity with non-filarial gut worm infections. 16 (19.75%) cats had reciprocal titers ranging from 320 to 2,560. Only 1 (1.23%) cat from this group was antigenemic.
    Matched MeSH terms: Elephantiasis, Filarial/immunology
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