Displaying publications 1 - 20 of 90 in total

Abstract:
Sort:
  1. Zarina Mohd Zawawi, Tengku Rogayah Tengku Abdul Rashid, Amir Hussien Adiee, Murni Maya Sari, Ravindran Thayan
    MyJurnal
    Introduction: Dengue virus (DENV), Zika virus (ZIKV) and Chikungunya virus (CHIKV) are Arboviruses that are transmitted by the same vector, Aedes aegypti. Dengue has become a global problem since the Second World War and is common in more than 110 countries. In Malaysia, dengue is a major disease burden as total economic costs to the country as a result of dengue is close to RM1.05 billion in 2010 and estimated to rise to 1.3 billion by 2020. Apart from Dengue, Zika and Chikungunya are the other important mosquito borne diseases in Malaysia. The aim of this study was to develop a multiplex real-time assay for simultaneous detection of DENV, ZIKV and CHIKV in clinical specimens. Methods: The published singleplex protocols were used with key modifications to implement a triplex assay. A one-step multiplex real-time RT-PCR assay was developed that can simultaneously detect RNA of DENV, ZIKV and CHIKV with good performance for a routine diagnostic use. The assay was evaluated for inter- and intra-reproducibility by mean CT value. The diagnostic sensitivity was tested with 135 archived samples which had been defined positive or negative by routine singleplex assays. Whole blood, plasma and urines were used in this study. Results: Intra- and inter-reproducibility and sensitivity varied from 0.10% to 4.73% and from 0.45% to 5.98% for each virus respectively. The specificity of detection was 100%. The multiplex real-time RT-PCR assay showed concordance with test results performed by routine singleplex assays. No cross reaction was observed for any of the clinical samples. Conclusion: The development of a rapid, sensitive and specific molecular assay for DENV, ZIKV and CHIKV infections will produce a greater diagnostic capacity in our laboratory. This multiplex approach is cost effective and robust with the concurrent detection of 3 viruses of public health concern.
    Matched MeSH terms: Chikungunya virus
  2. Zandi K
    Methods Mol Biol, 2016;1426:255-62.
    PMID: 27233278 DOI: 10.1007/978-1-4939-3618-2_23
    Screening of viral inhibitors through induction of cytopathic effects (CPE) by conventional method has been applied for various viruses including Chikungunya virus (CHIKV), a significant arbovirus. However, it does not provide the information about cytopathic effect from the beginning and throughout the course of virus replication. Conventionally, most of the approaches are constructed on laborious end-point assays which are not capable for detecting minute and rapid changes in cellular morphology. Therefore, we developed a label-free and dynamical method for monitoring the cellular features that comprises cell attachment, proliferation, and viral cytopathogenicity, known as the xCELLigence real-time cell analysis (RTCA). In this chapter, we provide a RTCA protocol for quantitative analysis of CHIKV replication using an infected Vero cell line treated with ribavirin as an in vitro model.
    Matched MeSH terms: Chikungunya virus/drug effects*
  3. Yusoff AF, Mustafa AN, Husaain HM, Hamzah WM, Yusof AM, Harun R, et al.
    BMC Infect Dis, 2013 May 08;13:211.
    PMID: 23656634 DOI: 10.1186/1471-2334-13-211
    BACKGROUND: The aims of the study were to assess the risk factors in relation to cross border activities, exposure to mosquito bite and preventive measures taken.An outbreak of chikungunya virus (CHIKV) infection in Malaysia has been reported in Klang, Selangor (1998) and Bagan Panchor, Perak (2006). In 2009, CHIKV infection re-emerged in some states in Malaysia. It raises the possibilities that re-emergence is part of the epidemics in neighbouring countries or the disease is endemic in Malaysia. For this reason, A community-based case control study was carried out in the state of Kelantan.

    METHODS: Prospective case finding was performed from June to December 2009. Those who presented with signs and symptoms of CHIKV infection were investigated. We designed a case control study to assess the risk factors. Assessment consisted of answering questions, undergoing a medical examination, and being tested for the presence of IgM antibodies to CHIKV. Descriptive epidemiological studies were conducted by reviewing both the national surveillance and laboratory data. Multivariable logistic regression analysis was performed to determine risk factors contributing to the illness. Cases were determined by positive to RT-PCR or serological for antibodies by IgM. CHIKV specificity was confirmed by DNA sequencing.

    RESULTS: There were 129 suspected cases and 176 controls. Among suspected cases, 54.4% were diagnosed to have CHIKV infection. Among the controls, 30.1% were found to be positive to serology for antibodies [IgM, 14.2% and IgG, 15.9%]. For analytic study and based on laboratory case definition, 95 were considered as cases and 123 as controls. Those who were positive to IgG were excluded. CHIKV infection affected all ages and mostly between 50-59 years old. Staying together in the same house with infected patients and working as rubber tappers were at a higher risk of infection. The usage of Mosquito coil insecticide had shown to be a significant protective factor. Most cases were treated as outpatient, only 7.5% needed hospitalization. The CHIKV infection was attributable to central/east African genotype CHIKV.

    CONCLUSIONS: In this study, cross border activity was not a significant risk factor although Thailand and Malaysia shared the same CHIKV genotype during the episode of infections.

    Matched MeSH terms: Chikungunya virus/genetics*; Chikungunya virus/isolation & purification
  4. Yap ML, Klose T, Urakami A, Hasan SS, Akahata W, Rossmann MG
    Proc Natl Acad Sci U S A, 2017 12 26;114(52):13703-13707.
    PMID: 29203665 DOI: 10.1073/pnas.1713166114
    Cleavage of the alphavirus precursor glycoprotein p62 into the E2 and E3 glycoproteins before assembly with the nucleocapsid is the key to producing fusion-competent mature spikes on alphaviruses. Here we present a cryo-EM, 6.8-Å resolution structure of an "immature" Chikungunya virus in which the cleavage site has been mutated to inhibit proteolysis. The spikes in the immature virus have a larger radius and are less compact than in the mature virus. Furthermore, domains B on the E2 glycoproteins have less freedom of movement in the immature virus, keeping the fusion loops protected under domain B. In addition, the nucleocapsid of the immature virus is more compact than in the mature virus, protecting a conserved ribosome-binding site in the capsid protein from exposure. These differences suggest that the posttranslational processing of the spikes and nucleocapsid is necessary to produce infectious virus.
    Matched MeSH terms: Chikungunya virus/metabolism; Chikungunya virus/ultrastructure*; Chikungunya virus/chemistry*
  5. Wong HV, Vythilingam I, Sulaiman WY, Lulla A, Merits A, Chan YF, et al.
    Am J Trop Med Hyg, 2016 Jan;94(1):182-6.
    PMID: 26598564 DOI: 10.4269/ajtmh.15-0318
    Vertical transmission may contribute to the maintenance of arthropod-borne viruses, but its existence in chikungunya virus (CHIKV) is unclear. Experimental vertical transmission of infectious clones of CHIKV in Aedes aegypti mosquitoes from Malaysia was investigated. Eggs and adult progeny from the second gonotrophic cycles of infected parental mosquitoes were tested. Using polymerase chain reaction (PCR), 56.3% of pooled eggs and 10% of adult progeny had detectable CHIKV RNA, but no samples had detectable infectious virus by plaque assay. Transfected CHIKV RNA from PCR-positive eggs did not yield infectious virus in BHK-21 cells. Thus, vertical transmission of viable CHIKV was not demonstrated. Noninfectious CHIKV RNA persists in eggs and progeny of infected Ae. aegypti, but the mechanism and significance are unknown. There is insufficient evidence to conclude that vertical transmission exists in CHIKV, as positive results reported in previous studies were almost exclusively based only on viral RNA detection.
    Matched MeSH terms: Chikungunya virus/genetics*; Chikungunya virus/isolation & purification*
  6. Wong HV, Chan YF, Sam IC, Sulaiman WY, Vythilingam I
    Methods Mol Biol, 2016;1426:119-28.
    PMID: 27233266 DOI: 10.1007/978-1-4939-3618-2_11
    In vivo infection of mosquitoes is an important method to study and characterize arthropod-borne viruses. Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that is transmitted primarily by Aedes mosquitoes. In this chapter, we describe a protocol for infection of CHIKV in two species of Aedes mosquitoes, Aedes aegypti and Aedes albopictus, together with the isolation of CHIKV in different parts of the infected mosquito such as midgut, legs, wings, salivary gland, head, and saliva. This allows the study of viral infection, replication and dissemination within the mosquito vector.
    Matched MeSH terms: Chikungunya virus/isolation & purification; Chikungunya virus/pathogenicity*; Chikungunya virus/physiology
  7. Wei Chiam C, Fun Chan Y, Chai Ong K, Thong Wong K, Sam IC
    J Gen Virol, 2015 Nov;96(11):3243-3254.
    PMID: 26276497 DOI: 10.1099/jgv.0.000263
    Chikungunya virus (CHIKV), an alphavirus of the family Togaviridae, causes fever, polyarthritis and rash. There are three genotypes: West African, Asian and East/Central/South African (ECSA). The latter two genotypes have caused global outbreaks in recent years. Recent ECSA CHIKV outbreaks have been associated with severe neurological disease, but it is not known if different CHIKV genotypes are associated with different neurovirulence. In this study, the neurovirulence of Asian (MY/06/37348) and ECSA (MY/08/065) strains of CHIKV isolated in Malaysia were compared. Intracerebral inoculation of either virus into suckling mice was followed by virus titration, histopathology and gene expression analysis of the harvested brains. Both strains of CHIKV replicated similarly, yet mice infected with MY/06/37348 showed higher mortality. Histopathology findings showed that both CHIKV strains spread within the brain (where CHIKV antigen was localized to astrocytes and neurons) and beyond to skeletal muscle. In MY/06/37348-infected mice, apoptosis, which is associated with neurovirulence in alphaviruses, was observed earlier in brains. Comparison of gene expression showed that a pro-apoptotic gene (eIF2αK2) was upregulated at higher levels in MY/06/37348-infected mice, while genes involved in anti-apoptosis (BIRC3), antiviral responses and central nervous system protection (including CD40, IL-10RA, MyD88 and PYCARD) were upregulated more highly in MY/08/065-infected mice. In conclusion, the higher mortality observed following MY/06/37348 infection in mice is due not to higher viral replication in the brain, but to differentially expressed genes involved in host immune responses. These findings may help to identify therapeutic strategies and biomarkers for neurological CHIKV infections.
    Matched MeSH terms: Chikungunya virus/classification; Chikungunya virus/genetics; Chikungunya virus/isolation & purification*; Chikungunya virus/pathogenicity*
  8. Wang SM, Ali UH, Sekaran SD, Thayan R
    Methods Mol Biol, 2016;1426:105-17.
    PMID: 27233265 DOI: 10.1007/978-1-4939-3618-2_10
    Real-time PCR assay has many advantages over conventional PCR methods, including rapidity, quantitative measurement, low risk of contamination, high sensitivity, high specificity, and ease of standardization (Mackay et al., Nucleic Acids Res 30:1292-1305, 2002). The real-time PCR system relies upon the measurement of a fluorescent reporter during PCR, in which the amount of emitted fluorescence is directly proportional to the amount of the PCR product in a reaction (Gibsons et al., Genome Res 6:995-1001, 1996). Here, we describe the use of SYBR Green I-based and TaqMan(®) real-time reverse transcription polymerase chain reaction (RT-PCR) for the detection and quantification of Chikungunya virus (CHIKV).
    Matched MeSH terms: Chikungunya virus/genetics*; Chikungunya virus/isolation & purification*
  9. Vythilingam I, Sam JI, Chan YF, Khaw LT, Sulaiman WY
    Front Microbiol, 2016;7:1452.
    PMID: 27679623 DOI: 10.3389/fmicb.2016.01452
    Zika virus (ZIKV) has now become a global public health concern. The vectors for ZIKV are Aedes aegypti and A. albopictus. Both these mosquitoes are predominant in Southeast Asia and are also responsible for the spread of other arboviral diseases like dengue virus and chikungunya virus. The incidence of dengue has been increasing over the years and this is of concern to public health workers. Simple laboratory tools for the detection of ZIKV is also lacking. In the absence of drugs and vaccine for these arboviral diseases, vector control is the main option for surveillance and control. Aedes larval surveys have been the hallmark of dengue control along with larviciding and fogging when cases are reported. However, we need new paradigms and options for control of these vectors. The current situation in Southeast Asia clearly proves that effective strategies for vector control need to be proactive and not reactive. This will be the way forward to control epidemics of these diseases inclusive of ZIKV until a vaccine becomes available.
    Matched MeSH terms: Chikungunya virus
  10. Viennet E, Knope K, Faddy HM, Williams CR, Harley D
    Commun Dis Intell Q Rep, 2013 Jun;37(2):E136-43.
    PMID: 24168087
    Chikungunya virus (CHIKV) is a major threat to Australia given the distribution of competent vectors, and the large number of travellers returning from endemic regions. We describe current knowledge of CHIKV importations into Australia, and quantify reported viraemic cases, with the aim of facilitating the formulation of public health policy and ensuring maintenance of blood safety.
    Matched MeSH terms: Chikungunya virus/isolation & purification*
  11. Ummul Haninah A, Vasan SS, Ravindran T, Chandru A, Lee HL, Shamala Devi S
    Trop Biomed, 2010 Dec;27(3):611-23.
    PMID: 21399603 MyJurnal
    This paper reports the development of a one-step SYBR-Green I-based realtime RT-PCR assay for the detection and quantification of Chikungunya virus (CHIKV) in human, monkey and mosquito samples by targeting the E1 structural gene. A preliminary evaluation of this assay has been successfully completed using 71 samples, consisting of a panel of negative control sera, sera from healthy individuals, sera from patients with acute disease from which CHIKV had been isolated, as well as monkey sera and adult mosquito samples obtained during the chikungunya fever outbreak in Malaysia in 2008. The assay was found to be 100-fold more sensitive than the conventional RT-PCR with a detection limit of 4.12x10(0) RNA copies/μl. The specificity of the assay was tested against other related viruses such as Dengue (serotypes 1-4), Japanese encephalitis, Herpes Simplex, Parainfluenza, Sindbis, Ross River, Yellow fever and West Nile viruses. The sensitivity, specificity and efficiency of this assay were 100%, 100% and 96.8% respectively. This study on early diagnostics is of importance to all endemic countries, especially Malaysia, which has been facing increasingly frequent and bigger outbreaks due to this virus since 1999.
    Matched MeSH terms: Chikungunya virus/isolation & purification*
  12. Tun MM, Thant KZ, Inoue S, Nabeshima T, Aoki K, Kyaw AK, et al.
    Emerg Infect Dis, 2014 Aug;20(8):1378-81.
    PMID: 25062511 DOI: 10.3201/eid2008.131431
    In 2010, chikungunya virus of the East Central South African genotype was isolated from 4 children in Myanmyar who had dengue-like symptoms. Phylogenetic analysis of the E1 gene revealed that the isolates were closely related to isolates from China, Thailand, and Malaysia that harbor the A226V mutation in this gene.
    Matched MeSH terms: Chikungunya virus/classification*; Chikungunya virus/genetics*
  13. Tuite AR, Watts AG, Khan K, Bogoch II
    Infect Dis Model, 2019;4:251-256.
    PMID: 31667444 DOI: 10.1016/j.idm.2019.09.001
    Southern Thailand has been experiencing a large chikungunya virus (CHIKV) outbreak since October 2018. Given the magnitude and duration of the outbreak and its location in a popular tourist destination, we sought to determine international case exportation risk and identify countries at greatest risk of receiving travel-associated imported CHIKV cases. We used a probabilistic model to estimate the expected number of exported cases from Southern Thailand between October 2018 and April 2019. The model incorporated data on CHIKV natural history, infection rates in Southern Thailand, average length of stay for tourists, and international outbound air passenger numbers from the outbreak area. For countries highly connected to Southern Thailand by air travel, we ran 1000 simulations to estimate the expected number of imported cases. We also identified destination countries with conditions suitable for autochthonous CHIKV transmission. Over the outbreak period, we estimated that an average of 125 (95% credible interval (CrI): 102-149) cases would be exported from Southern Thailand to international destinations via air travel. China was projected to receive the most cases (43, 95% CrI: 30-56), followed by Singapore (7, 95% CrI: 2-12) and Malaysia (5, 95% CrI: 1-10). Twenty-three countries were projected to receive at least one imported case, and 64% of these countries had one or more regions that could potentially support autochthonous CHIKV transmission. The overall risk of international exportation of CHIKV cases associated with the outbreak is Southern Thailand is high. Our model projections are consistent with recent reports of CHIKV in travelers returning from the region. Countries should be alert to the possibility of CHIKV infection in returning travelers, particularly in regions where autochthonous transmission is possible.
    Matched MeSH terms: Chikungunya virus
  14. Tongco AMP, Rivera WL
    Trop Biomed, 2023 Jun 01;40(2):129-137.
    PMID: 37650398 DOI: 10.47665/tb.40.2.002
    Chikungunya virus (CHIKV) is a neglected tropical pathogen that causes fever and long-lasting severe arthralgia. Despite its high morbidity, there is still no licensed specific therapeutic option for it. This study proposes a multi-epitope subunit vaccine candidate for CHIKV, designed using computational methods. It was based on the E2 spike glycoprotein in CHIKV, from which T- and B-cell epitopes were predicted and then refined. The pan HLA DR-binding epitope (PADRE) was added to this refined construct, then simulated compared with the native protein, where it was predicted to elicit more than twice the number of antibody titers. Thus, this construct is potentially effective against CHIKV, which further experimentation using live models would be able to verify. This study also demonstrates the feasibility of using rational tools in the future to further optimize vaccine design.
    Matched MeSH terms: Chikungunya virus*
  15. Thio CL, Yusof R, Ashrafzadeh A, Bahari S, Abdul-Rahman PS, Karsani SA
    PLoS One, 2015;10(6):e0129033.
    PMID: 26083627 DOI: 10.1371/journal.pone.0129033
    The Chikungunya virus (CHIKV) is an arthropod borne virus. In the last 50 years, it has been the cause of numerous outbreaks in tropical and temperate regions, worldwide. There is limited understanding regarding the underlying molecular mechanisms involved in CHIKV replication and how the virus interacts with its host. In the present study, comparative proteomics was used to identify secreted host proteins that changed in abundance in response to early CHIKV infection. Two-dimensional gel electrophoresis was used to analyse and compare the secretome profiles of WRL-68 cells infected with CHIKV against mock control WRL-68 cells. The analysis identified 25 regulated proteins in CHIKV infected cells. STRING network analysis was then used to predict biological processes that may be affected by these proteins. The processes predicted to be affected include signal transduction, cellular component and extracellular matrix (ECM) organization, regulation of cytokine stimulus and immune response. These results provide an initial view of CHIKV may affect the secretome of infected cells during early infection. The results presented here will compliment earlier results from the study of late host response. However, functional characterization will be necessary to further enhance our understanding of the roles played by these proteins in the early stages of CHIKV infection in humans.
    Matched MeSH terms: Chikungunya virus/physiology*
  16. Thayan R, Yusof MA, Saat Z, Sekaran SD, Wang SM
    Methods Mol Biol, 2016;1426:11-9.
    PMID: 27233257 DOI: 10.1007/978-1-4939-3618-2_2
    Molecular surveillance of Chikungunya virus (CHIKV) is important as it provides data on the circulating CHIKV genotypes in endemic countries and enabling activation of measures to be taken in the event of a pending outbreak. Molecular surveillance is carried out by first detecting CHIKV in susceptible humans or among field-caught mosquitoes. This is followed by sequencing a selected region of the virus which will provide evidence on the source of the virus and possible association of the virus to increased cases of Chikungunya infections.
    Matched MeSH terms: Chikungunya virus/classification; Chikungunya virus/genetics*; Chikungunya virus/isolation & purification
  17. Tesh RB, Gajdusek DC, Garruto RM, Cross JH, Rosen L
    Am J Trop Med Hyg, 1975 Jul;24(4):664-75.
    PMID: 1155702
    Plaque reduction neutralization tests, using five group A arboviruses (chikungunya, Ross River, Getah, Bebaru and Sindbis), were done on sera from human populations in 44 Southeast Asia and Pacific island localities. Specificity of the plaque neutralization test was determined by examining convalescent sera from patients with known alphavirus infections. Chikungunya-specific neutralizing antibodies were demonstrated in sera of persons living in South Vietnam, Northern Malaysia, Indonesia (Kalimantan and Sulawesi), as well as Luzon, Marinduque, Cebu and Mindanao islands in the Philippines. Evidence of Ross River virus infection was found among populations living in West New Guinea and Papua New Guinea mainland, the Bismark Archipelago, Rossel Island and the Solomon Islands. There appeared to be no geographic overlap in the distribution of chikungunya and Ross River viruses, with the separation in their distribution corresponding with Weber's line in the Pacific. Sindbis neutralizing antibodies were found in 7 of 21 populations sampled, but in general the prevalence of infection was low. Four sera, from Vietnam, Malaysia and Mindanao gave monospecific reactions with Getah virus. No evidence of specific Bebaru virus infection was detected. The epidemiology of these five alphaviruses in Southeast Asia and the Pacific islands is discussed.
    Matched MeSH terms: Chikungunya virus/immunology
  18. Tan KK, Sy AK, Tandoc AO, Khoo JJ, Sulaiman S, Chang LY, et al.
    Sci Rep, 2015 Jul 23;5:12279.
    PMID: 26201250 DOI: 10.1038/srep12279
    Outbreaks involving the Asian genotype Chikungunya virus (CHIKV) caused over one million infections in the Americas recently. The outbreak was preceded by a major nationwide outbreak in the Philippines. We examined the phylogenetic and phylogeographic relationships of representative CHIKV isolates obtained from the 2012 Philippines outbreak with other CHIKV isolates collected globally. Asian CHIKV isolated from the Philippines, China, Micronesia and Caribbean regions were found closely related, herein denoted as Cosmopolitan Asian CHIKV (CACV). Three adaptive amino acid substitutions in nsP3 (D483N), E1 (P397L) and E3 (Q19R) were identified among CACV. Acquisition of the nsP3-483N mutation in Compostela Valley followed by E1-397L/E3-19R in Laguna preceded the nationwide spread in the Philippines. The China isolates possessed two of the amino acid substitutions, nsP3-D483N and E1-P397L whereas the Micronesian and Caribbean CHIKV inherited all the three amino acid substitutions. The unique amino acid substitutions observed among the isolates suggest multiple independent virus dissemination events. The possible biological importance of the specific genetic signatures associated with the rapid global of the virus is not known and warrant future in-depth study and epidemiological follow-up. Molecular evidence, however, supports the Philippines outbreak as the possible origin of the CACV.
    Matched MeSH terms: Chikungunya virus/classification; Chikungunya virus/genetics*; Chikungunya virus/isolation & purification
  19. Syuhadaratul Aini Mohamat, Nor Fazila Che Mat, Najmo Ibrahim Barkhadle2, Tuan Nur Akmalina Mat Jusoh, Rafidah Hanim Shueb
    MyJurnal
    Chikungunya is an infection caused by chikungunya virus which at present has spread to new countries and con- tinents. Chikungunya is associated with self-limiting and non-fatal infection in the past. However, in recent times, increased severity of the disease has been reported resulting in health and economic burden. The threat and bur- den of chikungunya would grow in future in the absence of specific antiviral or vaccine to control or eliminate the infection. This review discusses chikungunya in general including transmission of its etiological agent and clinical manifestations of the disease. Subsequently, management and treatment of chikungunya virus will be reviewed with particular emphasis on natural products or their active compounds with potential anti-chikungunya virus activities.
    Matched MeSH terms: Chikungunya virus
  20. Sum, Magdline Sia Henry, Andrew, Anna, Maling, Milda Aren
    MyJurnal
    Chikungunya is an acute febrile illness caused by chikungunya virus (CHIKV). In this study, the envelope E1 gene of CHIKV was cloned and expressed in a baculovirus system. The recombinant E1 protein with N-term 6-His residues protein was successfully expressed and purified as confirmed by SDS-PAGE and western blot analysis. The seroreactivity of the recombinant protein was evaluated in immunoassay for anti-CHIKV IgM and IgG antibodies. The recombinant antigen showed 69% sensitivity and 100% specificity for anti-CHIKV IgG by dot blot assay. Detection of anti-CHIKV IgM by dot assay showed 79% sensitivity and 100% specificity. No cross reactivity of the antigen was observed with anti-dengue virus serum samples. The results strongly support that the recombinant E1 protein has potential to be used as diagnostic antigen. The used of the antigen in a dot blot assay gives an advantage for laboratory detection without the need of any specialised equipment.
    Matched MeSH terms: Chikungunya virus
Filters
Contact Us

Please provide feedback to Administrator (afdal@afpm.org.my)

External Links