Displaying publications 1 - 20 of 34 in total

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  1. Benacer D, Mohd Zain SN, Amran F, Galloway RL, Thong KL
    Am J Trop Med Hyg, 2013 Apr;88(4):704-9.
    PMID: 23358635 DOI: 10.4269/ajtmh.12-0662
    Rats are considered the principal maintenance hosts of Leptospira. The objectives of this study were isolation and identification of Leptospira serovars circulating among urban rat populations in Kuala Lumpur. Three hundred urban rats (73% Rattus rattus and 27% R. norvegicus) from three different sites were trapped. Twenty cultures were positive for Leptospira using dark-field microscopy. R. rattus was the dominant carrier (70%). Polymerase chain reaction (PCR) confirmed that all isolates were pathogenic Leptospira species. Two Leptospira serogroups, Javanica and Bataviae, were identified using microscopic agglutination test (MAT). Pulsed-field gel electrophoresis (PFGE) identified two serovars in the urban rat populations: L. borgpetersenii serovar Javanica (85%) and L. interrogans serovar Bataviae (15%). We conclude that these two serovars are the major serovars circulating among the urban rat populations in Kuala Lumpur. Despite the low infection rate reported, the high pathogenicity of these serovars raises concern of public health risks caused by rodent transmission of leptospirosis.
  2. Benacer D, Woh PY, Mohd Zain SN, Amran F, Thong KL
    Microbes Environ, 2013;28(1):135-40.
    PMID: 23363618
    Leptospira species were studied in water and soils from selected urban sites in Malaysia. A total of 151 water (n=121) and soil (n=30) samples were collected from 12 recreational lakes and wet markets. All samples were filtered and inoculated into semi-solid Ellinghausen and McCullough modified by Johnson and Harris (EMJH) media supplemented with additional 5-fluorouracil. The cultures were then incubated at 30°C and observed under a dark field microscope with intervals of 10 days. A PCR assay targeting the rrs gene was used to confirm the genus Leptospira among the isolates. Subsequently, the pathogenic status of the isolates was determined using primer sets G1/G2 and Sapro1/Sapro2, which target the secY and rrs genes, respectively. The isolates were identified at serogroup level using the microscopic agglutination test (MAT) while their genetic diversity was assessed by pulsed field gel electrophoresis (PFGE). Based on dark field microscopy, 23.1% (28/121) water and 23.3% (7/30) soil cultures were positive for Leptospira spp. Of the 35 positive cultures, only 8 were pure and confirmed as Leptospira genus by PCR assay. Two out of 8 isolates were confirmed as pathogenic, 5 were saprophytic and one was intermediate. These 8 isolates were negative for the 25 reference hyperimmune rabbit sera tested in the MAT. PFGE showed that all 8 of these environmental Leptospira spp. were genetically diverse. In conclusion, the presence of pathogenic Leptospira spp. in the urban Malaysian environment may indicate and highlight the importance of water screening, especially in recreational lakes, in order to minimize any chance of Leptospira infection.
  3. Mohd Tap R, Kamarudin NA, Ginsapu SJ, Ahmed Bakri AR, Ahmad N, Amran F, et al.
    Genome Announc, 2018 Apr 05;6(14).
    PMID: 29622608 DOI: 10.1128/genomeA.00166-18
    Candida pseudohaemulonii is phylogenetically close to the C. haemulonii complex and exhibits resistance to amphotericin B and azole agents. We report here the draft genome sequence of C. pseudohaemulonii UZ153_17 isolated from the blood culture of a neutropenic patient. The draft genome is 3,532,003,666 bp in length, with 579,838 reads, 130 contigs, and a G+C content of 47.15%.
  4. Vincent AT, Schiettekatte O, Goarant C, Neela VK, Bernet E, Thibeaux R, et al.
    PLoS Negl Trop Dis, 2019 05;13(5):e0007270.
    PMID: 31120895 DOI: 10.1371/journal.pntd.0007270
    The causative agents of leptospirosis are responsible for an emerging zoonotic disease worldwide. One of the major routes of transmission for leptospirosis is the natural environment contaminated with the urine of a wide range of reservoir animals. Soils and surface waters also host a high diversity of non-pathogenic Leptospira and species for which the virulence status is not clearly established. The genus Leptospira is currently divided into 35 species classified into three phylogenetic clusters, which supposedly correlate with the virulence of the bacteria. In this study, a total of 90 Leptospira strains isolated from different environments worldwide including Japan, Malaysia, New Caledonia, Algeria, mainland France, and the island of Mayotte in the Indian Ocean were sequenced. A comparison of average nucleotide identity (ANI) values of genomes of the 90 isolates and representative genomes of known species revealed 30 new Leptospira species. These data also supported the existence of two clades and 4 subclades. To avoid classification that strongly implies assumption on the virulence status of the lineages, we called them P1, P2, S1, S2. One of these subclades has not yet been described and is composed of Leptospira idonii and 4 novel species that are phylogenetically related to the saprophytes. We then investigated genome diversity and evolutionary relationships among members of the genus Leptospira by studying the pangenome and core gene sets. Our data enable the identification of genome features, genes and domains that are important for each subclade, thereby laying the foundation for refining the classification of this complex bacterial genus. We also shed light on atypical genomic features of a group of species that includes the species often associated with human infection, suggesting a specific and ongoing evolution of this group of species that will require more attention. In conclusion, we have uncovered a massive species diversity and revealed a novel subclade in environmental samples collected worldwide and we have redefined the classification of species in the genus. The implication of several new potentially infectious Leptospira species for human and animal health remains to be determined but our data also provide new insights into the emergence of virulence in the pathogenic species.
  5. Amran F, Mohd Khalid MK, Mohamad S, Mat Ripen A, Ahmad N, Goris MG, et al.
    Genome Announc, 2016;4(5).
    PMID: 27609924 DOI: 10.1128/genomeA.00956-16
    Leptospira interrogans serovar Bataviae was recently identified as one of the persistent Leptospira serovars in Malaysia. Here, we report the draft genome sequence of the L. interrogans serovar Bataviae strain LepIMR 22 isolated from kidney of a rodent in Johor, Malaysia.
  6. Arushothy R, Amran F, Samsuddin N, Ahmad N, Nathan S
    PLoS Negl Trop Dis, 2020 12;14(12):e0008979.
    PMID: 33370273 DOI: 10.1371/journal.pntd.0008979
    BACKGROUND: Melioidosis is a neglected tropical disease with rising global public health and clinical importance. Melioidosis is endemic in Southeast Asia and Northern Australia and is of increasing concern in Malaysia. Despite a number of reported studies from Malaysia, these reports are limited to certain parts of the country and do not provide a cohesive link between epidemiology of melioidosis cases and the nation-wide distribution of the causative agent Burkholderia pseudomallei.

    METHODOLOGY/PRINCIPLE FINDINGS: Here we report on the distribution of B. pseudomallei sequence types (STs) in Malaysia and how the STs are related to STs globally. We obtained 84 culture-confirmed B. pseudomallei from confirmed septicaemic melioidosis patients from all over Malaysia. Prior to performing Multi Locus Sequence Typing, the isolates were subjected to antimicrobial susceptibility testing and detection of the YLF/BTFC genes and BimA allele. Up to 90.5% of the isolates were sensitive to all antimicrobials tested while resistance was observed for antimicrobials typically administered during the eradication stage of treatment. YLF gene cluster and bimABp allele variant were detected in all the isolates. The epidemiological distribution patterns of the Malaysian B. pseudomallei isolates were analysed in silico using phylogenetic tools and compared to Southeast Asian and world-wide isolates. Genotyping of the 84 Malaysian B. pseudomallei isolates revealed 29 different STs of which 6 (7.1%) were novel. ST50 was identified as the group founder followed by subgroup founders ST376, ST211 and ST84. A low-level diversity is noted for the B. pseudomallei isolates described in this study while phylogenetic analysis associated the Malaysian STs to Southeast Asian isolates especially isolates from Thailand. Further analysis also showed a strong association that implicates agriculture and domestication activities as high-risk routes of infection.

    CONCLUSIONS/SIGNIFICANCE: In conclusion, MLST analysis of B. pseudomallei clinical isolates from all states in Malaysia revealed low diversity and a close association to Southeast Asian isolates.

  7. Rao M, Amran F, Kamaruzaman AA, Hakim Esa HA, Abdul Hameed A, Mohamed Shabery NA
    Am J Trop Med Hyg, 2021 01;104(1):216-218.
    PMID: 33289472 DOI: 10.4269/ajtmh.20-0267
    Leptospirosis is an important zoonotic disease with worldwide distribution and nonspecific clinical manifestation. We report a case of fatal leptospirosis in a previously healthy woman with a causative agent. A young adult Indian woman was brought in dead to the forensic department. Ten days before, she developed fever, dizziness with headache, myalgia, diarrhea, and vomiting. Routine inquest and autopsy were performed on the deceased, revealing hemorrhagic lungs with extensive intra-alveolar hemorrhages, pale liver with dissociation and separation of hepatocyte plates, and edematous brain with histiocyte and lymphocyte infiltration in the parenchyma and meninges. Heart tissue depicts myocarditis and pericarditis inflammatory changes. Cerebrospinal fluid (CSF) was turbid in appearance with mildly elevated leukocytes, predominantly lymphocytes. Real-time PCR targeting lipL32 gene of pathogenic Leptospira was detected in the blood, CSF, brain, kidney, heart, and liver. The genetic profile of the causative agent was ST149 (multi-locus sequence typing Scheme 3). This study illustrates the usefulness of Leptospira PCR assay in postmortem diagnosis and addresses the need for further surveillance to identify the epidemiological link of the disease.
  8. Mohd Ali MR, Sum JS, Aminuddin Baki NN, Choong YS, Nor Amdan NA, Amran F, et al.
    Int J Biol Macromol, 2021 Jan 31;168:289-300.
    PMID: 33310091 DOI: 10.1016/j.ijbiomac.2020.12.062
    Leptospirosis is a potentially fatal zoonosis that is caused by spirochete Leptospira. The signs and symptoms of leptospirosis are usually varied, allowing it to be mistaken for other causes of acute febrile syndromes. Thus, early diagnosis and identification of a specific agent in clinical samples is crucial for effective treatment. This study was aimed to develop specific monoclonal antibodies against LipL21 antigen for future use in leptospirosis rapid and accurate immunoassay. A recombinant LipL21 (rLipL21) antigen was optimized for expression and evaluated for immunogenicity. Then, a naïve phage antibody library was utilized to identify single chain fragment variable (scFv) clones against the rLipL21 antigen. A total of 47 clones were analysed through monoclonal phage ELISA. However, after taking into consideration the background OD405 values, only 4 clones were sent for sequencing to determine human germline sequences. The sequence analysis showed that all 4 clones are identical. The in silico analysis of scFv-lip-1 complex indicated that the charged residues of scFv CDRs are responsible for the recognition with rLipL21 epitopes. The generated monoclonal antibody against rLipL21 will be evaluated as a detection reagent for the diagnosis of human leptospirosis in a future study.
  9. Riazi M, Zainul FZ, Bahaman AR, Amran F, Khalilpour A
    Indian J Med Res, 2014 Feb;139(2):308-13.
    PMID: 24718408
    Leptospirosis is a widespread zoonotic disease and a public health problem, particularly in tropical and subtropical countries. Varied clinical manifestations of the disease frequently lead to misdiagnosis resulting in life-threatening multi-organ complications. Therefore, early laboratory investigation using an appropriate diagnostic approach is crucial. In the present study, a potential protein marker was identified and evaluated for its usefulness in the serodiagnosis of acute leptospirosis.
  10. Daud AB, Mohd Fuzi NMH, Wan Mohammad WMZ, Amran F, Ismail N, Arshad MM, et al.
    Int J Occup Environ Med, 2018 04;9(2):88-96.
    PMID: 29667646 DOI: 10.15171/ijoem.2018.1164
    BACKGROUND: Leptospirosis is an emerging zoonosis and its occurrence has been reported to be rising globally. The environment plays an important role in the survival of Leptospira and determines the risk of infection. Those who were exposed to and had contact with contaminated environment through their occupational, recreational and other activities can be infected with the organism.

    OBJECTIVE: To determine the seroprevalence of leptospirosis among cattle farmers, prevalence of pathogenic Leptospira, and the workplace environmental risk factors for leptospirosis among cattle farmers in northeastern Malaysia.

    METHODS: A cross-sectional study involving 120 cattle farmers was conducted. The participants answered an interviewer-guided questionnaire that consisted of sociodemographic and workplace environment characteristics questionnaire, before having their blood sample taken for microscopic agglutination test (MAT). Seropositivity was determined using a cut-off titer of ≥1:100. 248 environmental samples were also collected from the cattle farms for polymerase chain reaction (PCR).

    RESULTS: The overall seroprevalence of leptospiral antibodies was 72.5% (95% CI 63.5% to 80.1%) and the prevalence of pathogenic Leptospira in the cattle farms environment was 12.1% (95% CI 8.4% to 17.0%). The independent factors associated with seropositivity of leptospirosis among cattle farmers were positive pathogenic Leptospira in the environment (Adj OR 5.90, 95% CI 1.34 to 26.01) and presence of garbage dumping in the farm (Adj OR 2.40, 95% CI 1.02 to 5.65).

    CONCLUSION: Preventing leptospirosis incidence among cattle farmers necessitates changes in work environment. Identifying modifiable factors may also contribute to the reduction of infection.

  11. Daud A, Fuzi NMHM, Arshad MM, Kamarudin S, Mohammad WMZW, Amran F, et al.
    Vet World, 2018 Jun;11(6):840-844.
    PMID: 30034179 DOI: 10.14202/vetworld.2018.840-844
    Background: Leptospirosis is a zoonotic disease that infects human and livestock which causes economic losses to the farmers. It has been reported as one of the causes of reproductive failure in cattle and other ruminants, determining abortions, stillbirth, weak newborns, and decrease in their growth rate and milk production.

    Aim: The objectives of this study were to determine the leptospirosis seroprevalence and to identify the predominant infecting serovars among cattle.

    Materials and Methods: A cross-sectional study involving 420 cattle from six randomly selected districts in Kelantan was conducted. A serological test using the microscopic agglutination test was conducted in the Institute of Medical Research with a cutoff titer for seropositivity of ≥1:100.

    Results: The overall prevalence of leptospirosis seropositivity among cattle in this study was 81.7% (95% confidence interval: 63.5, 80.1). The most common reaction obtained with the sera tested was from the serovar Sarawak with 78.8%.

    Conclusion: A high seroprevalence of leptospiral antibodies was found among cattle in Northeastern Malaysia. These findings urge that more studies are required to determine the reasons for the high seroprevalence among the cattle along with its transmission and pathogenicity of the local serovar Sarawak.

  12. Amran F, Aziz MN, Ibrahim HM, Atiqah NH, Parameswari S, Hafiza MR, et al.
    J Med Microbiol, 2011 Sep;60(Pt 9):1312-1316.
    PMID: 21459913 DOI: 10.1099/jmm.0.027631-0
    The in vitro antifungal susceptibilities of 159 clinical isolates of Candida species from patients with invasive candidiasis in Kuala Lumpur Hospital, Malaysia, were determined against amphotericin B, fluconazole, voriconazole, itraconazole and caspofungin. The most common species were Candida albicans (71 isolates), Candida parapsilosis (42 isolates), Candida tropicalis (27 isolates) and Candida glabrata (12 isolates). The susceptibility tests were carried out using an E-test. The MIC breakpoints were based on Clinical Laboratory Standards Institute criteria. Amphotericin B and voriconazole showed the best activities against all the isolates tested, with MIC(90) values of ≤1 µg ml(-1) for all major species. Only one Candida lusitaniae isolate was resistant to amphotericin B, and all the isolates were susceptible to voriconazole. In total, six isolates were resistant to fluconazole, comprising two isolates of C. albicans, two of C. parapsilosis, one C. tropicalis and one C. glabrata, and all of these isolates showed cross-resistance to itraconazole. The MIC(90) of itraconazole was highest for C. glabrata and C. parapsilosis. Caspofungin was active against most of the isolates except for five isolates of C. parapsilosis. The MIC(90) of caspofungin against C. parapsilosis was 3 µg ml(-1). In conclusion, amphotericin B remains the most active antifungal agent against most Candida species except for C. lusitaniae. Voriconazole is the best alternative for fluconazole- or itraconizole-resistant isolates. Although five of the C. parapsilosis isolates showed in vitro resistance to caspofungin, more clinical correlation studies need to be carried out to confirm the significance of these findings. Currently, despite the increase in usage of antifungals in our hospitals, especially in the management of febrile neutropenia patients, the antifungal-resistance problem among clinically important Candida isolates in Kuala Lumpur Hospital is not yet worrying. However, continued antifungal-susceptibility surveillance needs to be conducted to monitor the antifungal-susceptibility trends of Candida species and other opportunistic fungal pathogens.
  13. Shamsusah NA, Agustar HK, Amran F, Hod R
    Microbiol Resour Announc, 2021 Sep 30;10(39):e0072321.
    PMID: 34591676 DOI: 10.1128/MRA.00723-21
    Previously, a novel Leptospira strain (BJ3) was isolated from the soil of an ex situ wild animal conservation area in Perak, Malaysia. Molecular identification via whole-genome sequencing confirmed that the strain was Leptospira yasudae. Here, we report the draft genome sequence of L. yasudae strain BJ3.
  14. Amran F, Liow YL, Halim NAN
    J Korean Med Sci, 2018 Apr 23;33(17):e131.
    PMID: 29686599 DOI: 10.3346/jkms.2018.33.e131
    Leptospirosis is a febrile zoonotic disease. Routine diagnosis of leptospirosis is based on the detection of specific antibodies with serological tests. The aim of our study was to determine the usefulness of immunochromatographic assay (ICA), ImmuneMed Leptospira IgM Duo Rapid test kit from Korea, in rapid screening of acute leptospirosis in emergency cases with limited expertise. A total of 197 serum samples (93 positive, 104 negative) were selected randomly. The test has good diagnostic sensitivity 73% and specificity 90%. With positive predictive value of 87% and negative predictive value of 79%, this reassures patients have higher chance of correct diagnosis. This ICA is acceptable for screening of leptospirosis but confirmation with microscopic agglutination test should follow.
  15. Arushothy R, Ahmad N, Amran F, Hashim R, Samsuddin N, Che Azih CR
    Genome Announc, 2018 Apr 19;6(16).
    PMID: 29674530 DOI: 10.1128/genomeA.00167-18
    After the introduction of the pneumococcal conjugate vaccine in Malaysia in recent years, the emergence of nonvaccine serotypes is of concern, particularly the antibiotic-resistant strains, with an increase specifically in serotype 15A. Here, we report the draft genome sequence of Streptococcus pneumoniae strain SS40_16, isolated from the blood sample of a 19-month-old female in 2016. SS40_16 is a multidrug-resistant strain with resistance to penicillin (MIC, ≥2 µg/ml), tetracycline, and trimethoprim-sulfamethoxazole. The strain belongs to serotype 15A and sequence type 1591 (ST1591).
  16. Arushothy R, Ahmad N, Amran F, Hashim R, Samsudin N, Azih CRC
    Int J Infect Dis, 2019 Mar;80:129-133.
    PMID: 30572022 DOI: 10.1016/j.ijid.2018.12.009
    OBJECTIVE: This study was performed to analyze the serotype distribution of Streptococcus pneumoniae causing invasive pneumococcal disease (IPD) in children aged 5 years and under in Malaysia and to assess the antimicrobial resistance.

    METHODS: From 2014 to 2017, a total of 245 invasive S. pneumoniae isolates from children ≤5 years of age were received from hospitals all around Malaysia. All isolates were identified and subjected to serotyping and antimicrobial susceptibility testing.

    RESULTS: Of the 245 isolates, 117 (48.0%) were from children aged <1year, 46 (19.05%) were from children aged 1-2 years, and 82 (33.0%) were from children aged 2-5 years. The most common serotypes were 14 (26.9%), 6B (19.6%), 19A (11.8%), 6A (10.6%), and 19F (6.9%) and vaccine coverage was 88.2% for PCV13, 64.1% for PCV10, and 63.3% for PCV7. Resistance to penicillin was 0.2% for non-meningitis cases and 22.2% for meningitis cases; erythromycin resistance was reported in 42.9%, co-trimoxazole in 35.9%, and tetracycline in 42.9%.

    CONCLUSIONS: Serotypes 14, 6B, 19A, 6A, and 19F were the most common serotypes isolated from children with IPD in Malaysia during this pre-vaccination era. The lack of reports on the serotype distribution has limited action for the implementation of PCV in the national immunization programme (NIP). The information from this study may benefit future policies for the introduction of PCV in the Malaysian NIP and ultimately may reduce the morbidity and mortality among children in Malaysia.

  17. Rao M, Amran F, Aqilla N
    Can J Infect Dis Med Microbiol, 2019;2019:5763595.
    PMID: 30881530 DOI: 10.1155/2019/5763595
    Introduction: Leptospirosis is an acute febrile illness, known for its protean clinical manifestations and the challenge in differentiating from other infectious diseases. Standardized confirmatory test is antibody dependent and not accessible by the suburban community. This study measures efficiency of an immune-chromatographic assay, Leptocheck WB, in detecting acute leptospirosis.

    Methods: A total of 142 sera were used for kit evaluation. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated by comparing rapid kit results with gold standard laboratory, microscopic agglutination test (MAT).

    Results: We found this rapid kit to have a sensitivity and specificity of 66.6% and 78.9%, respectively, whereas the PPV and NPV of the kit appeared to be 73.3% and 73.2%, respectively.

    Discussion: Test efficiency of this rapid kit is reasonable. It is specific in detecting leptospiral antibody and assures clinician of accurate diagnosis by having higher PPV and NPV. It is prompt and efficient in comparison with conventional methods in assisting differential diagnosis. High sensitivity and specificity leptospirosis rapid test is indeed a crucial measure to assist the diagnosis of acute undifferentiated febrile illnesses.

  18. Othman N, Intan HI, Yip CW, Alias M, Amran F
    J Trop Pediatr, 2007 Feb;53(1):55-8.
    PMID: 17237115
    We report a case of an 8-year-old aborigine boy referred to our hospital for respiratory insufficiency with skin eruptions over the trunk and limbs. The skin condition was diagnosed as acquired ichthyosis. He also had a non-bleeding form of disseminated intravascular coagulopathy. Radiograph of the lungs showed bilateral perihilar opacities with bilateral pleural effusion. The diagnosis of leptospirosis was confirmed by a 4-fold rise in microagglutinating titre and polymerase chain reaction assay.
  19. Othman N, Yip CW, Intan HI, Zainuddin Z, Amran F
    Ann Trop Paediatr, 2006 Sep;26(3):259-62.
    PMID: 16925966
    A 7-year-old boy, referred with lymphoma, presented with prolonged fever and intra-abdominal lymphadenopathy demonstrated on computed tomography (CT) of the abdomen. Blood culture isolated Penicillium marneffei. The patient was subsequently proven serologically to be positive for human immunodeficiency virus (HIV). Treatment with amphotericin B followed by itraconazole was successful. A high level of clinical suspicion and awareness is necessary for early diagnosis of penicilliosis, especially in an era of an increasing prevalence of HIV in this region.
  20. Hii SYF, Ali NA, Ahmad N, Amran F
    J Med Microbiol, 2017 Nov;66(11):1623-1627.
    PMID: 29048275 DOI: 10.1099/jmm.0.000611
    Melioidosis is an endemic infectious disease in Southeast Asia and northern Australia, caused by Burkholderia pseudomallei. However, the incidence rate in Malaysia is not well documented. The high mortality rate and broad range of clinical presentations require rapid and accurate diagnosis for appropriate treatment. This study compared the efficacy of in-house IgM and IgG ELISA methods using a local B. pseudomallei strain. The diagnostic accuracy of the in-house IgG ELISA was better than that of the IgM ELISA: sensitivity (IgG: 84.71 %, IgM: 76.14 %) and specificity (IgG: 93.64 %, IgM: 90.17 %); positive predictive value (IgG: 86.75 %, IgM: 79.76 %) and negative predictive value (IgG: 92.57 %, IgM: 89.66 %); likelihood ratio (LR) [IgG: 13.32, IgM: 7.75 (LR+); IgG: 0.16, IgM: 0.26 (LR-)], and was supported by the observation of the absorbance value in comparisons between culture and serology sampling. In-house IgG ELISA was shown to be useful as an early diagnostic tool for melioidosis.
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