Serological surveys have been widely used in South-East Asia to determine the presence and activity of arboviruses. The haemagglutination-inhibition test has been most frequently employed but complement-fixation and neutralization tests have also been used in some investigations.Although virus isolations provide the most conclusive evidence, they can be carried out in a few specialized centres only, and serological surveys are very important for studying the distribution of arboviruses.The surveys have shown that group B arboviruses (principally all four types of dengue, Japanese encephalitis, and West Nile) are widely prevalent. Dengue and Japanese encephalitis viruses are more widespread than West Nile virus, which was not known previously to extend east of India although recent survyes have shown that its range extends to Burma. Japanese encephalitis is frequent in most of South-East Asia but in India is found mainly in eastern and south-eastern parts of the country. Kyasanur Forest disease (KFD) and Langat viruses are the only tick-borne group B arboviruses definitely known to occur in the region, the former in India, the latter in Malaysia. KFD virus has been isolated only from a small focus in Mysore, although human and animal sera containing neutralizing antibodies to this virus have been found sporadically in widely scattered areas. Among the group A arboviruses, chikungunya and Sindbis have been detected in serological surveys, but the former has not yet been found in Malaysia.
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.
A survey of the activity of three alphaviruses (Sindbis, getah and chikungunya) in Peninsular Malaysia was conducted between 1962 and 1970. Serum samples were examined from 3,917 vertebrates representing a wide variety of wild and domestic animals throughout the peninsula for hemagglutination-inhibiting and neutralizing antibodies. A total of 548,939 mosquitoes were collected from different habitats, including jungle, rural, suburban and urban areas, and the majority of the females taken were examined for the presence of virus. Two strains of Sindbis virus and one strain of getah virus were isolated from pools of Culex mosquitoes collected in and around domestic animal shelters. Analysis of the serological results indicated that, 1) getah virus is associated principally with large domestic animals, particularly swine, 2) Sindbis virus is associated with large domestic animals and birds, especially domestic ducks, and 3) chikungunya virus, which has not yet been isolated in Malaysia, appeared to be present at a very low level of activity, probably with wild monkeys as the vertebrate hosts.
A serum survey of several characteristic groups of humans in urban, rural, and forested areas of Peninsular Malaysia for evidence of infection with three alphaviruses (Sindbis, getah, and chikungunya) was made on 4384 specimens collected between 1965 and 1969. Analysis of the serological results indicated that 1) persons residing in predominantly rural and forested areas have higher frequencies of specific alphavirus antibody of all three viruses than persons residing in urban areas, 2) human infection with chikungunya virus appears to be at a low level of activity but is widespread, although more common and recent in the northern part of the country, and 3) Sindbis and getah viruses probably do not represent a threat to the public health, but chikungunya virus remains a potential menance and may be responsible for future epidemics transmitted by A. aegypti and A. albopictus mosquitoes.
Many countries neighboring Malaysia have reported human infections by chikungunya virus, a mosquito-borne togavirus belonging to the genus Alphavirus. However, although there is serological evidence of its presence in Malaysia, chikungunya virus has not been known to be associated with clinical illness in the country. An outbreak of chikungunya virus occurred in Klang, Malaysia, between December 1998 and February 1999. The majority of the cases were in adults and the clinical presentation was similar to classical chikungunya infections. Malaysia is heavily dependent on migrant workers from countries where chikungunya is endemic. It is speculated that the virus has been re-introduced into the country through the movement of these workers.
A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein E1 (E1) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354- and 294-base pair (bp) cDNA fragments were amplified effectively from the nsP1 and E1 genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial E1 gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype.
Emerging and re-emerging infectious diseases have become a major global problem. Malaysia appears to be an epicenter for such infections and in recent years, several outbreaks have occurred resulting in loss of lives and economic hardships. In this paper, we discussed the outbreaks of leptospirosis, enterovirus 71 encephalitis, chikungunya polyarthritis and Nipah encephalitis and how a developing country such as Malaysia managed the situation with the help of international agencies and organisations. Many valuable lessons were learned and by sharing our experience, it is hoped that we will be in a better position to handle future outbreaks and prevent their spread to countries in the region.
A study of chikungunya virus was carried out to establish Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) as a rapid detection technique of the virus. The susceptibility of lab-colonized Aedes aegypti to chikungunya virus was also determined. Artificial membrane feeding technique was used to orally feed the mosquitoes with a human isolate of chikungunya virus. A total of 100 fully engorged female Ae. aegypti were obtained and maintained for 7 days. Seventy of them survived and then pooled at 10 individuals per pool. Total RNA was extracted from the samples and RT-PCR amplifications were carried out. Five out of 7 pools showed positive PCR band at 350-bp, indicating Ae. aegypti is a potential vector of chikungunya virus. The minimum infection rate (MIR) was 71% within these laboratory colonies. RT-PCR is a sensitive technique that is useful in detecting infected mosquitoes in epidemic areas. This technique can de used as a rapid detection method and provide an early virologic surveillance systems of chikungunya virus infected mosquitoes.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus which causes epidemic fever, rash and polyarthralgia in Africa and Asia. Two outbreaks have been reported in Malaysia, in Klang, Selangor (1998) and Bagan Panchor, Perak (2006). It is not known if the outbreaks were caused by the recent introduction of CHIKV, or if the virus was already circulating in Malaysia. Seroprevalence studies from the 1960s suggested previous disease activity in certain parts of the country. In Asia, CHIKV is thought to be transmitted by the same mosquitoes as dengue, Aedes aegypti and Ae. albopictus. Due to similarities in clinical presentation with dengue, limited awareness, and a lack of laboratory diagnostic capability, CHIKV is probably often underdiagnosed or misdiagnosed as dengue. Treatment is supportive. The prognosis is generally good, although some patients experience chronic arthritis. With no vaccine or antiviral available, prevention and control depends on surveillance, early identification of outbreaks, and vector control. CHIKV should be borne in mind in sporadic cases, and in patients epidemiologically linked to ongoing local or international outbreaks or endemic areas.
An outbreak of Chikugunya (CHIK) fever occurred among the fishing community in Bagan Pancor, Perak. The outbreak was laboratory confirmed within 48 hours after the receipt of the specimens. Fifty-three patients' serum samples were submitted for laboratory investigation and 47 (88.7%) were confirmed to be positive for CHIK infection by RT-PCR, and/or virus isolation, and/or in-house immunoflourescent test. RT-PCR and virus isolation were the tests of choice for patients with illness of four days or less and detection of CHIK specific IgM for those with more than four days of fever. The nucleic acid sequence based on the 354- and 294-bp of the nsP1 and E1 genes of the CHIK virus detected from pools of adults Aedes aegypti mosquitoes were identical to those CHIKV virus isolated from humans in the same locality. Phylogenetic analysis of the CHIK virus based on the 257 nts partial E1 gene indicates that Bagan Panchor's strain was closely related to the first CHIK virus isolated during the outbreak in Klang in 1998.
Chikungunya virus infection recently reemerged in Malaysia after 7 years of nondetection. Genomic sequences of recovered isolates were highly similar to those of Malaysian isolates from the 1998 outbreak. The reemergence of the infection is not part of the epidemics in other Indian Ocean countries but raises the possibility that chikungunya virus is endemic in Malaysia.
Since its isolation in Tanzania in 1953, chikungunya virus has caused periodic outbreaks in both tropical Africa and Asia. In the last decade, the virus has shown not only increased activity but has expanded its geographical locations, thus classical delineation of various genotypes of chikungunya virus to specific geographic locales no longer holds true. Rapid mass movement of people and the constant presence of the right vectors in this region could have contributed to the change in virus ecology. This paper documents the first detection of chikungunya virus of Central/East genotype in Malaysia from a patient who was most likely infected with the virus during her visit to India. Without good Aedes vector measures, only time will tell whether this genotype rather than the existing endemic genotype will subsequently cause the next chikungunya outbreak in Malaysia.
Chikungunya is an acute febrile illness caused by an alphavirus which is transmitted by infective Aedes mosquitoes. Two previous outbreaks of chikungunya in Malaysia were due to chikungunya virus of Asian genotype. The present outbreak involved two adjoining areas in the suburb of Ipoh city within the Kinta district of Perak, a state in the northern part of Peninsular Malaysia. Thirty seven residents in the main outbreak area and two patients in the secondary area were laboratory confirmed to be infected with the virus. The index case was a 44-year Indian man who visited Paramakudi, Tamil Naidu, India on 21st November 2006 and returned home on 30th of November 2006, and subsequently developed high fever and joint pain on the 3rd of December 2006. A number of chikungunya virus isolates were isolated from both patients and Aedes albopictus mosquitoes in the affected areas. Molecular study showed that the chikungunya virus causing the Kinta outbreak was of the Central/East African genotype which occurred for the first time in Malaysia.
During an outbreak of chikungunya in a dengue hyperendemic area within the Kinta district of Perak, two patients with acute febrile illness were laboratory confirmed to have co-infection of both dengue and chikungunya viruses in their blood. The concomitant presence of two types of viruses transmitted by the same vector in a susceptible population contributed to the resultant event. A good understanding of virus vector ecology in association with population dynamics and wider application of improved laboratory techniques by using different cell-lines suited for optimal replication of each type of virus and the correct utilization of powerful molecular techniques will enhance accurate diagnosis of these infectious diseases.
Many triggering factors for onset of emerging infectious diseases are now recognised, such as: globalisation, demographic increase, population movements, international trade, urbanisation, forest destruction, climate changes, loss in biodiversity, and extreme life conditions such as poverty, famine and war. Epidemic burden is often leading to disasters, in terms of human losses, as well as economic, political or social consequences. These outbreaks may jeopardize within a few weeks or months, industry, trade, or tourism. While dengue and its most severe forms (hemorrhagic and shock syndrome) is spreading all over the tropical world, another arbovirosis, chikungunya disease dramatically spread in Indian Ocean islands where 30 to 75% of population were infected in 2005 and 2006, and then extended its progression towards India, Sri Lanka, Indonesia, Malaysia, Maldives islands with more than a million people infected with the East-African strain, replacing the former Asian strain which was known to prevail more than 30 years ago in India. Patients experience sequelae with disability, work loss, and rarely severe outcome recently identified in La Réunion and Mayotte (French overseas territories). No country, no part of the world may consider itself as protected against such events. However, consequences of emerging or re-emerging diseases are more and more unacceptable when they impact the poorest countries of the world. Viruses, bacteria, as well as wild animals, birds, or arthropods are not stopped by borders. It is time now to promote barriers against infectious diseases, including prevention, anticipation, disease surveillance and research. This is not only for humanitarian reasons, but also for contributing to a sustainable development with equity for worldwide population. This report presents comprehensive actions taken in 2006 for tracing the epidemic and mobilise research, as requested to the task force set up by the Prime Minister by March 20, 2006.
In this study, artificial membrane feeding technique was used to orally feed Aedes aegypti with dengue and chikungunya viruses. Virus detection was carried out by reverse transcriptase polymerase chain reaction. The study did not detect dual infection of Ae. aegypti with dengue and chikungunya virus from the same pool or from individual mosquitoes. Oral receptivity of Ae. aegypti to chikungunya virus was higher than that of dengue virus.
In 2008, an outbreak of chikungunya infection occurred in Johor. We performed a retrospective review of all laboratory confirmed adult chikungunya cases admitted to Hospital Sultanah Aminah, Johor Bahru from April to August 2008, looking into clinical and laboratory features. A total of 18 laboratory confirmed cases of chikungunya were identified with patients presenting with fever, joint pain, rash and vomiting. Haemorrhagic signs were not seen. Lymphopenia, neutropenia, thrombocytopenia, raised liver enzymes and deranged coagulation profile were the prominent laboratory findings. We hope this study can help guide physician making a diagnosis of chikungunya against other arborviruses infection.
BACKGROUND: Chikungunya virus (CHIKV) of the Central/East African genotype has caused large outbreaks worldwide in recent years. In Malaysia, limited CHIKV outbreaks of the endemic Asian and imported Central/East African genotypes were reported in 1998 and 2006. Since April 2008, an unprecedented nationwide outbreak has affected Malaysia.
OBJECTIVE: To study the molecular epidemiology of the current Malaysian CHIKV outbreak, and to evaluate cross-neutralisation activity of serum from infected patients against isolates of Asian and Central/East African genotypes.
STUDY DESIGN: Serum samples were collected from 83 patients presenting in 2008, and tested with PCR for the E1 gene, virus isolation, and for IgM. Phylogenetic analysis was performed on partial E1 gene sequences of 837bp length. Convalescent serum from the current outbreak and Bagan Panchor outbreak (Asian genotype, 2006) were tested for cross-neutralising activity against representative strains from each outbreak.
RESULTS: CHIKV was confirmed in 34 patients (41.0%). The current outbreak strain has the A226V mutation in the E1 structural protein, and grouped with Central/East African isolates from recent global outbreaks. Serum cross-neutralisation activity against both Central/East African and Asian genotypes was observed at titres from 40 to 1280.
CONCLUSIONS: The CHIKV strain causing the largest Malaysian outbreak is of the Central/East African genotype. The presence of the A226V mutation, which enhances transmissibility of CHIKV by Aedes albopictus, may explain the extensive spread especially in rural areas. Serum cross-neutralisation of different genotypes may aid potential vaccines and limit the effect of future outbreaks.
Chikungunya infections were detected in Singapore among returning travelers who had visited friends and relatives (VFR) in India and Malaysia. These sporadic imported cases occurred over a year before the 2008 chikungunya outbreaks in Singapore, demonstrating the potential for introducing this emerging viral infection into new areas via VFR travel.