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  1. Yeng C, Osman E, Mohamed Z, Noordin R
    Electrophoresis, 2010 Dec;31(23-24):3843-9.
    PMID: 21080484 DOI: 10.1002/elps.201000038
    Toxoplasma gondii infection in pregnant women may result in abortion and foetal abnormalities, and may be life-threatening in immunocompromised hosts. To identify the potential infection markers of this disease, 2-DE and Western blot methods were employed to study the parasite circulating antigens and host-specific proteins in the sera of T. gondii-infected individuals. The comparisons were made between serum protein profiles of infected (n=31) and normal (n=10) subjects. Antigenic proteins were identified by immunoblotting using pooled sera and monoclonal anti-human IgM-HRP. Selected protein spots were characterised using mass spectrometry. Prominent differences were observed when serum samples of T. gondii-infected individuals and normal controls were compared. A significant up-regulation of host-specific proteins, α(2)-HS glycoprotein and α(1)-B glycoprotein, was also observed in the silver-stained gels of both active and chronic infections. However, only α(2)-HS glycoprotein and α(1)-B glycoprotein in the active infection showed immunoreactivity in Western blots. In addition, three spots of T. gondii proteins were detected, namely (i) hypothetical protein chrXII: 3984434-3 TGME 49, (ii) dual specificity protein phosphatase, catalytic domain TGME 49 and (iii) NADPH-cytochrome p450 reductase TGME 49. Thus, 2-DE approach followed by Western blotting has enabled the identification of five potential infection markers for the diagnosis of toxoplasmosis: three are parasite-specific proteins and two are host-specific proteins.
  2. Ahmad N., Osman, E., Abdul Ghani, N.A., Leong, W.Y., Arzaee, M.Z., Seow, Y.Y., et al.
    Medicine & Health, 2020;15(2):108-123.
    MyJurnal
    Toksoplasmosis pendam dapat menyebabkan pelbagai gangguan hormon dan tingkah laku dalam hos terjangkit. Kami berhasrat untuk mengkaji sero-prevalens Toxoplasma gondii (T. gondii) pendam serta hubungan antara jangkitan dengan pengetahuan dan tingkah laku dalam kalangan 400 ibu hamil. Sampel plasma diuji untuk kehadiran antibodi IgG T. gondii dan soal selidik berstruktur digunakan untuk merekodkan ciri-ciri sosio-demografi responden, maklumat umum dan pengetahuan mengenai faktor risiko, gejala, masa jangkitan, pengetahuan pencegahan serta tingkah laku pencegahan toksoplasmosis. Sero-prevalensi toksoplasmosis pendam dalam wanita hamil adalah 31.8%. Kajian menunjukkan, 69.5% daripada mereka mempunyai kurang pengetahuan mengenai toksoplasmosis. Walau bagaimanapun, majoritinya (99.8%) mengamalkan tingkah laku pencegahan. Analisis regresi logistik berganda menunjukkan wanita hamil dengan tahap pendidikan rendah mempunyai hampir dua kali lebih risiko (nisbah ods terlaras: 1.91, 95% SK 1.18, 3.10; p = 0.008) untuk T. gondii IgG seropositif. Wanita hamil yang mempunyai sejarah perubatan lalu mempunyai dua kali lebih kemungkinan (nisbah ods terlaras: 2.32, 95% SK 1.32, 4.06; p = 0.003) untuk T. gondii IgG seropositif. Selain itu, wanita yang tidak pasti mengenai mod penyebaran penyakit melalui pemindahan darah mempunyai empat kali lebih ods (nisbah ods terlaras: 3.93, 95% SK 1.54, 10.01; p = 0.004) untuk sero-prevalens kronik toksoplasmosis. Wanita yang tidak pasti mengenai keperluan menghindari kucing liar mempunyai nisbah ods terlaras: 0.42 (95% SK 0.24, 0.71, p = 0.001) untuk sero-prevalens kronik toksoplasmosis. Penterjemahan pengetahuan tentang toksoplasmosis kepada amalan tingkah laku pencegahan melalui program pendidikan kesihatan adalah penting untuk mengurangkan risiko penularan penyakit ini dalam kalangan wanita hamil.
  3. Nimir AR, Osman E, Ibrahim IA, Saliem AM
    BMJ Case Rep, 2013;2013.
    PMID: 23580678 DOI: 10.1136/bcr-2013-008803
    A 31-year-old Malaysian man was presented with an episode of seizures by the roadside, after having been recently diagnosed as HIV positive accompanied with miliary tuberculosis. On physical examination, he was oriented to person, but not to time or place. There was no neck stiffness or papilloedema. The other systemic examination was unremarkable. Chest examination revealed crepitations at the upper zone of the right lung. After diagnosis suspicion, the case was confirmed as toxoplasma encephalitis by MRI and serological tests. Patient was treated with trimethoprim/sulfamethoxazole 480-2400 mg/day with folinic acid supplement for 60 days. Two months later, a repeat brain MRI showed resolution of the cerebral lesions.
  4. Mohamed Yusoff PS, Osman E, Raja Sabudin RZA
    Malays J Pathol, 2021 Apr;43(1):9-18.
    PMID: 33903300
    Disseminated microsporidiosis is a life-threatening disease resulting from the haematogenous spread of microsporidia species. The diagnosis is challenging owing to its subtle nonspecific clinical presentation, which usually reflects the underlying organ involved. Therefore, a high index of suspicion is required for early diagnosis. Besides, tools for confirmatory laboratory diagnosis are limited. Currently, there is no direct diagnostic method that can detect the infection without involving invasive procedures. Clinical confirmation of disseminated microsporidiosis is usually based on light and transmission electron microscopy of infected tissue specimens. These are then followed by species detection using polymerase chain reaction (PCR). Disseminated microsporidiosis shows the potential to be cleared up by albendazole or fumagillin if they are detected and treated early. Based on a series of case reports, this review aims to present a current update on disseminated microsporidiosis with emphasis on the clinical manifestations based on the organ system infected, diagnostic approach and treatment of this devastating condition.
  5. Saadatnia G, Mohamed Z, Ghaffarifar F, Osman E, Moghadam ZK, Noordin R
    APMIS, 2012 Jan;120(1):47-55.
    PMID: 22151308 DOI: 10.1111/j.1600-0463.2011.02810.x
    Infection with Toxoplasma gondii is widespread and important in humans, especially pregnant women and immunosuppressed patients. A panel of tests is usually required for diagnosis toxoplasmosis. Excretory secretory antigen (ESA) is highly immunogenic, and thus it is a good candidate for investigation into new infection markers. ESA was prepared from tachyzoites of RH strain of T. gondii by mice intraperitoneal infection. Sera were obtained from several categories of individuals who differed in their status of anti-Toxoplasma IgM, IgG and IgG avidity antibodies. The ESA was subjected to SDS-PAGE, two-dimensional gel electrophoresis and Western blot analysis. Antigenic bands of approximate molecular weights of 12, 20 and 30 kDa, when probed with anti-human IgM-HRP and IgA-HRP, showed good potential as infection markers. The highest sensitivity of the bands was 98.7% with combination of IgM and IgA blots with sera of patients with anti-Toxoplasma IgM+ IgG+. The specificities were 84% and 70% with sera from other infections and healthy controls in IgM blots and IgA blots respectively. By mass spectrometry, the 12 kDa protein was identified as thioredoxin. The two top proteins identified for 20 kDa molecule were microneme protein 10 and dense granule protein 7; whereas that for 30 kDa were phosphoglycerate mutase 1 and phosphoglycerate mutase.
  6. Anuar TS, Bakar NH, Al-Mekhlafi HM, Moktar N, Osman E
    PMID: 27405127
    The epidemiology and environmental factors affecting transmission of human microsporidiosis are poorly understood. We conducted the present study to determine the prevalence and risk factors associated with asymptomatic intestinal microsporidiosis among aboriginal school children in the Kuala Krau District, Pahang State, Malaysia. We collected stool samples from 255 school children and examined the samples using Gram-chromotrope Kinyoun stain. We also collected demographic, socioeconomic, environmental and personal hygiene information using a pre-tested questionnaire. Sixty-nine of the children was positive for microsporidia: 72.5% and 27.5% were low (1+) and moderate (2+) excretions of microsporidia spores, respectively. Univariate and multivariate analyses showed being aged 10 years (p = 0.026), using an unsafe water supply as a source for drinking water (p = 0.044) and having close contact with domestic animals (p = 0.031) were all significantly associated with microsporidial infection among study subjects. Our findings suggest asymptomatic intestinal microsporidiosis is common in the study population, more than previously reported. In the study population, control measures need to be implemented, such as good personal hygiene, proper sanitation and safe drinking water supply.
  7. Wahid W, Zahariluddin ASM, Kadir ZS, Sharip S, Idris ZM, Osman E
    Iran J Parasitol, 2021 10 12;16(3):512-517.
    PMID: 34630598 DOI: 10.18502/ijpa.v16i3.7106
    Schizophrenia is a serious mental disorder characterized by chronic relapsing episodes of psychosis. The disease is multifactorial, where infections, genetic vulnerability and environmental factors are involved in the development of the illness. Toxoplasma gondii is one of the parasites that has long been known associated with schizophrenia in many studies. To date, there is growing evidence of association between T. gondii infections and schizophrenia. Herein we report a rare case of reactivated toxoplasmosis in a schizophrenia individual. This patient was incidentally diagnosed with reactivated T. gondii infection. He denied any symptoms of toxoplasmosis but experienced a mild psychiatric auditory hallucination. Serology test for T. gondii immunoglobulin antibodies measured a high positive IgG titer (135.9 IU/ml) and negative for IgM. Interestingly, nested PCR exhibited a positive result for the type I strain of T. gondii dense granular (GRA) 7 gene (GRA7). This case highlights the detection of probable reactivation of toxoplasmosis in an immunocompetent schizophrenic patient without psychiatric treatment-resistant and remains asymptomatic for toxoplasmosis. Both serology and molecular tools have been a helpful aid in establishing the diagnosis. Nonetheless, early detection as in this case may aid the patient management in the future.
  8. Noordin R, Osman E, Kalantari N, Anuar NS, Gorgani-Firouzjaee T, Sithithaworn P, et al.
    Acta Trop, 2022 Feb;226:106251.
    PMID: 34808116 DOI: 10.1016/j.actatropica.2021.106251
    Strongyloides stercoralis is a parasite that causes strongyloidiasis worldwide. It may lead to a life-long infection in immunocompetent people and hyperinfection in immunosuppressed patients. A point-of-care (POC) rapid test is helpful for patient diagnosis in resource-limited settings and as a detection tool in elimination/control programs. Previously, we reported a rapid IgG4 dipstick test (Ss Rapid®) for Strongyloides suitable for a laboratory setting. A POC cassette format of the test, which is field-applicable, has since been developed. Here, we report on a laboratory-based evaluation of the Ss Rapid® cas sette test on 285 sera. We assessed the diagnostic sensitivity of the Ss Rapid® cas sette with 32 sera, comprising samples from larval and/or DNA positive individuals from three countries. Additionally, we also tested samples from 33 seropositive endemic areas residents. We evaluated the diagnostic specificity of the test using 220 samples, comprising sera from other infections (n = 101), allergy cases with high IgE antibodies (n = 4), and blood donors (n = 115). The test showed high diagnostic sensitivity (97%, 31/32), and all sera of the seropositive endemic residents were reactive. It also showed high diagnostic specificity (94.5%, 208/220), and all false-positive samples tested negative after sera adsorption using recombinant NIE-coated microsphere beads. Additionally, we showed that the test worked with spiked whole blood samples. The study results showed that the SsRapid® cas sette test merits further laboratory and field evaluations.
  9. Noordin R, Osman E, Anuar NS, Juri NM, Rahumatullah A, Ahmad Hilmi NA
    Am J Trop Med Hyg, 2021 08 30;105(5):1214-1217.
    PMID: 34460427 DOI: 10.4269/ajtmh.21-0674
    A lateral flow rapid test for strongyloidiasis will greatly facilitate the control and elimination of the disease. Previously SsRapid prototype rapid test showed high diagnostic specificity to detect Strongyloides infection, determined using non-Strongyloides sera negative by IgG-ELISAs. Since high specificity is crucial before a test is used for public health control activities, further validation of its specificity is needed. Also, it needs to be ascertained whether non-Strongyloides sera positive by IgG-ELISAs and SsRapid are truly positive for Strongyloides or are cases of cross-reactivity. We performed 84 rapid tests (two types of dipsticks and cassettes) using 34 serum samples. They were divided into four groups based on Strongyloides infection and coinfection with other parasites and the availability of recombinant proteins and rapid tests for the latter. Sera was adsorbed using polystyrene microspheres beads separately coated with four recombinant parasite proteins. The small sample size is a limitation of this study; however, the overall results showed that the sera adsorption procedure was successful, and the SsRapid test is specific.
  10. Nor Azmi NJ, Mohamad S, Shahidan WNS, Taib H, Mohamed Z, Osman E
    Saudi Dent J, 2024 Feb;36(2):258-261.
    PMID: 38419993 DOI: 10.1016/j.sdentj.2023.11.014
    INTRODUCTION: Periodontal disease is the inflammation of the periodontium tissues surrounding the teeth, potentially leading to loss of tooth attachment. In individuals with periodontal disease, the presence of Trichomonas tenax, a parasitic protozoan of the oral cavity has been observed and its frequency tends to rise as the disease progresses.

    METHODS: A literature search was conducted in the online databases of PubMed, Google Scholar, Web of Science, and Scopus using the combination of keywords: "Trichomonas tenax" AND "periodontal disease" OR "gum disease", OR "oral disease" OR "periodontitis". A total of 9 articles satisfied the inclusion criteria and were included in this study.

    RESULTS: This review highlights the incidence of T. tenax with periodontal diseases, the risk factors that contribute to the infection of T. tenax and available detection methods for the identification of the protozoan.

    CONCLUSION: The inhabitation of the oral cavity by T. tenax prospers with the severity of periodontal diseases. Extensive research should be conducted to fully understand the potential pathogenic role and damaging effect of T. tenax in the oral cavity.

  11. Mohamed Yusoff PS, Arifin N, Periyasamy P, Tumian NR, Ismail F, Raja Sabudin RZA, et al.
    Malays J Pathol, 2024 Aug;46(2):299-306.
    PMID: 39207007
    Diagnosis of extraintestinal microsporidiosis is always hampered due to non-specific symptoms and difficulty in diagnosis. This study aimed to compare the diagnostic utility of blood and faecal-based polymerase chain reaction (PCR) to detect microsporidiosis in immunocompromised patients. A total of 42 immunocompromised patients consisting of HIV-infected and chemotherapy-treated patients were enrolled. Paired faecal and blood samples were collected and subjected to PCR to detect Enterocytozoon bieneusi and Encephalitozoon spp. Faecal samples were microscopically screened for microsporidia spores. Overall, 42.9% (18/42) of patients were positive for microsporidiosis. Of this, 19.0% (8/42) and 4.8% (2/42) were positive by blood and stool PCR respectively. Meanwhile, 33.3% (14/42) of the faecal specimens were microscopically positive. Among the positive patients, 22.2% (4/18) had microsporidia confirmed by blood PCR and stool microscopy, suggestive of dissemination. Interestingly, the stool specimen in which microsporidia spores were detected via microscopy is not positive via PCR method. This highlights the limitation of the faecal-based detection method and the important use of blood samples for diagnosing extraintestinal microsporidiosis. Only E. bieneusi species were detected in all PCR-positive samples. This study highlights the diagnostic value of blood PCR in diagnosing extraintestinal microsporidiosis infections.
  12. Shahrul Anuar T, M Al-Mekhlafi H, Abdul Ghani MK, Osman E, Mohd Yasin A, Nordin A, et al.
    PLoS One, 2012;7(10):e48165.
    PMID: 23133561 DOI: 10.1371/journal.pone.0048165
    BACKGROUND: Entamoeba histolytica/Entamoeba dispar/Entamoeba moshkovskii infection is still prevalent in rural Malaysia especially among Orang Asli communities. Currently, information on prevalence of this infection among different ethnic groups of Orang Asli is unavailable in Malaysia. To contribute to a better comprehension of the epidemiology of this infection, a cross-sectional study aimed at providing the first documented data on the prevalence and risk factors associated with E. histolytica/E. dispar/E. moshkovskii infection was carried out among three Orang Asli ethnic groups (Proto-Malay, Negrito, and Senoi) in selected villages in Negeri Sembilan, Perak, and Pahang states, Malaysia.

    METHODS/FINDINGS: Faecal samples were examined by formalin-ether sedimentation and trichrome staining techniques. Of 500 individuals, 8.7% (13/150) of Proto-Malay, 29.5% (41/139) of Negrito, and 18.5% (39/211) of Senoi were positive for E. histolytica/E. dispar/E. moshkovskii, respectively. The prevalence of this infection showed an age-dependency relationship, with higher rates observed among those aged less than 15 years in all ethnic groups studied. Multivariate analysis confirmed that not washing hands after playing with soils or gardening and presence of other family members infected with E. histolytica/E. dispar/E. moshkovskii were significant risk factors of infection among all ethnic groups. However, eating with hands, the consumption of raw vegetables, and close contact with domestic animals were identified as significant risk factors in Senoi.

    CONCLUSIONS: Essentially, the findings highlighted that E. histolytica/E. dispar/E. moshkovskii parasites are still prevalent in Malaysia. Further studies using molecular approaches to distinguish the morphologically identical species of pathogenic, E. histolytica from the non-pathogenic, E. dispar and E. moshkovskii are needed. The establishment of such data will be beneficial for the public health authorities in the planning and implementation of specific prevention and control strategies of this infection in different Orang Asli ethnic groups in Malaysia.

  13. Anuar TS, Al-Mekhlafi HM, Ghani MK, Osman E, Yasin AM, Nordin A, et al.
    Int J Parasitol, 2012 Aug;42(9):871-80.
    PMID: 22846786 DOI: 10.1016/j.ijpara.2012.07.003
    The flagellate protozoan parasite, Giardia intestinalis, is widely distributed throughout the world with a high prevalence in developing countries in the tropics and subtropics, including Malaysia. Approximately 200 million people are infected with the parasite globally, with 500,000 new cases reported annually. This cross-sectional study was conducted among three tribes of Orang Asli communities in Selangor, Perak and Pahang states of Malaysia. The main objective was to determine the prevalence of and risk factors for giardiasis. Stool samples were collected from 500 individuals aged between 2 and 74 years (males=219, females=281). The samples were examined with formalin-ether sedimentation and trichrome staining techniques. Socioeconomic data were collected through a pre-tested questionnaire. The overall prevalence of giardiasis was 20.0% with the highest prevalence in the Proto-Malays (33.3%) followed by Negritos (20.1%) and Senois (10.4%). The positive cases showed a decrease with increasing age and most of the positive cases were observed in individuals less than 24 years old. Males had significantly higher prevalence than females (χ(2)=5.283, P=0.022). Logistic regression analysis of the overall population studied and the Senoi tribe confirmed that being a child aged less than 15 years, being male, the consumption of raw vegetables and the presence of other family members infected with G. intestinalis were the main risk factors for giardiasis. The presence of other family members infected with G. intestinalis was the only risk factor highlighted in the Proto-Malay and Negrito tribes. Diarrhoea was significantly associated with giardiasis. However, the cause and effect relationship has yet to be determined. Thus, screening family members and treating the infected individuals are the main strategies that should be adopted by the public health authority in combating this infection in Orang Asli communities as well as health education regarding good personal and food hygiene practises.
  14. Munajat MB, Rahim MAFA, Wahid W, Seri Rakna MIM, Divis PCS, Chuangchaiya S, et al.
    Malar J, 2021 Apr 27;20(1):202.
    PMID: 33906645 DOI: 10.1186/s12936-021-03741-y
    BACKGROUND: Malaysia is on track towards malaria elimination. However, several cases of malaria still occur in the country. Contributing factors and communal aspects have noteworthy effects on any malaria elimination activities. Thus, assessing the community's knowledge, attitudes and practices (KAP) towards malaria is essential. This study was performed to evaluate KAP regarding malaria among the indigenous people (i.e. Orang Asli) in Peninsular Malaysia.

    METHODS: A household-based cross-sectional study was conducted in five remote villages (clusters) of Orang Asli located in the State of Kelantan, a central region of the country. Community members aged six years and above were interviewed. Demographic, socio-economic and KAP data on malaria were collected using a structured questionnaire and analysed using descriptive statistics.

    RESULTS: Overall, 536 individuals from 208 households were interviewed. Household indoor residual spraying (IRS) coverage and bed net ownership were 100% and 89.2%, respectively. A majority of respondents used mosquito bed nets every night (95.1%), but only 50.2% were aware that bed nets were used to prevent malaria. Nevertheless, almost all of the respondents (97.9%) were aware that malaria is transmitted by mosquitoes. Regarding practice for managing malaria, the most common practice adopted by the respondents was seeking treatment at the health facilities (70.9%), followed by self-purchase of medication from a local shop (12.7%), seeking treatment from a traditional healer (10.5%) and self-healing (5.9%). Concerning potential zoonotic malaria, about half of the respondents (47.2%) reported seeing monkeys from their houses and 20.1% reported entering nearby forests within the last 6 months.

    CONCLUSION: This study found that most populations living in the villages have an acceptable level of knowledge and awareness about malaria. However, positive attitudes and practices concerning managing malaria require marked improvement.

  15. Osman E, Amin NA, Noon TPM, Lahat SNH, Rosli MS, Sham SF, et al.
    Am J Trop Med Hyg, 2022 Jul 25;107(3):636-9.
    PMID: 35895335 DOI: 10.4269/ajtmh.22-0076
    Strongyloides infection may develop into fatal hyperinfection and dissemination syndrome in immunocompromised hosts. Despite suboptimal specificity issues, the detection of IgG antibodies by ELISA has been central in the serodiagnosis of Strongyloides infection. Recently, an IgG4-based lateral-flow test (SsRapid) using recombinant NIE (rNIE) protein with good diagnostic performance has been reported. This study assessed the result concordance between a commercial IgG-ELISA and the SsRapid. Additionally, we determined the Strongyloides seroprevalence and its association with clinical manifestations. Immunocompromised patients (N = 200) were from Hospital Canselor Tuanku Muhriz, Kuala Lumpur, Malaysia, and were diagnosed with HIV/AIDS, hematological malignancy, and solid organ cancers. Their plasma samples were tested using a commercial IgG-ELISA and SsRapid. A fair concordance (κ = 0.27-0.33; P < 0.05) among the tests was demonstrated. The SsRapid exhibited a significantly higher (P < 0.05) seroprevalence (10.5% [21/200]) compared with IgG-ELISA (7.5% [15/200]). After adsorption with rNIE, all SsRapid-positive samples tested negative with the rapid test, thus showing binding specificity. There was no significant association with clinical manifestations. This study revealed that SsRapid is a useful diagnostic tool for Strongyloides infection, and there is a notable seroprevalence among the immunocompromised patients.
  16. Dian ND, Muhammad AB, Azman EN, Eddie NA, Azmi NI, Yee VCT, et al.
    Am J Trop Med Hyg, 2023 Nov 01;109(5):1081-1085.
    PMID: 37748768 DOI: 10.4269/ajtmh.23-0184
    Malaysia has maintained zero cases of indigenous human malaria since 2018. However, zoonotic malaria is still prevalent in underdeveloped areas and hard-to-reach populations. This study aimed to determine the prevalence of malaria among remote indigenous communities in Peninsular Malaysia. A cross-sectional survey was conducted in six settlements in Kelantan state, from June to October 2019. Blood samples were tested for malaria using microscopy and nested polymerase chain reaction (nPCR) targeting the Plasmodium cytochrome c oxidase subunit III (cox3) gene. Of the 1,954 individuals who appeared healthy, no malaria parasites were found using microscopy. However, nPCR revealed seven cases of Plasmodium knowlesi mono-infection (0.4%), and six out of seven infections were in the group of 19 to 40 years old (P = 0.026). No human malaria species were detected by nPCR. Analysis of the DNA sequences also showed high similarity that reflects common ancestry to other P. knowlesi isolates. These findings indicate low submicroscopic P. knowlesi infections among indigenous communities in Malaysia, requiring PCR-based surveillance to support malaria control activities in the country.
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