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).
Introduction: Malaria is a public health concern and still endemic in Malaysia particularly in Sabah. Malaria is due to plasmodium parasite which can be further divided into human malaria and zoonotic malaria. Plasmodium knowlesi, a zoonotic malaria species, is increasing in trend and alarming despite of ongoing prevention and control measures especially in Sabah. Methods: This is a case-control study involving a total of 169 respondents which comprises of 43 cases and 126 controls from Mukim Paginatan and Mukim Nalapak, Ranau. A simple random sampling were used to select subjects. Results: This study found male, forest related and doing outdoor activity has higher risk of contracting malaria. Village house is another significant risk factor. The specific risk factors with the strongest association with malaria is the presence of monkey near housing or farm area (OR5.667; 95% CI: 2.429, 13.220), live near forest (OR 5.330; 95% CI: 1.209, 23.495), presence of pond or pool or river near house (OR 4.222; 95% CI: 1.552, 11.490), doing outdoor activity after 5 PM (OR 3.434; 95% CI: 1.522, 7.748) and ever overnight in the hut (OR 2.724; 95% CI: 1.334, 5.561). Conclusion: This study shows several risk factors that associated with malaria knowlesi. Based on the identified risk factors, preventive and control measure can be improved and optimized.
Introduction: Pertussis is known to cause infection and reinfection to everyone irrespective of ages and countries. Therefore, adults do require vaccination for protection against pertussis infection especially the HCW. However, the pertussis vaccine coverage is low among HCW due to low-risk perception. Protection Motivation Theory (PMT) is one of the most cited theories to explain risk perception and intention to change. Therefore, we developed a questionnaire based on the subconstructs of the PMT to assess the acceptance of the pertussis vaccine amongst the HCWs in Sabah and Sarawak. The motive of this study is to validate this questionnaire to see its validity and reli- ability. Method: Data was collected using a self-administered questionnaire via an online survey (Monkey Survey). The questionnaire was given to 250 HCW. Items that were identified as a problem were modified to increase reli- ability. Further validation was done among 853 HCWs working in various parts of Sabah and Sarawak. Results: The Cronbach alpha of the overall construct of PMT during the first pilot study was 0.66 and improved to 0.82. Principal components factor analysis using varimax rotations showed that the first four factors explained 28%, 2%, 9% and 5% of the variance respectively. Both the one level and two-level modelling indicated that it’s a good fit model. Conclu- sion: The study instrument that was developed for the study has been tested and proven to be relevant to assess the risk perception of an HCW towards pertussis.
A large focus of human infections with Plasmodium knowlesi, a simian parasite naturally found in long-tailed and pig-tailed macaques was discovered in the Kapit Division of Sarawak, Malaysian Borneo. A study was initiated to identify the vectors of malaria, to elucidate where transmission is taking place and to understand the bionomics of the vectors in Kapit.
Malaria is one of the most devastating infectious diseases of humans. It is problematic clinically and economically as it prevails in poorer countries and regions, strongly hindering socioeconomic development. The causative agents of malaria are unicellular protozoan parasites belonging to the genus Plasmodium. These parasites infect not only humans but also other vertebrates, from reptiles and birds to mammals. To date, over 200 species of Plasmodium have been formally described, and each species infects a certain range of hosts. Plasmodium species that naturally infect humans and cause malaria in large areas of the world are limited to five-P. falciparum, P. vivax, P. malariae, P. ovale and P. knowlesi. The first four are specific for humans, while P. knowlesi is naturally maintained in macaque monkeys and causes zoonotic malaria widely in South East Asia. Transmission of Plasmodium species between vertebrate hosts depends on an insect vector, which is usually the mosquito. The vector is not just a carrier but the definitive host, where sexual reproduction of Plasmodium species occurs, and the parasite's development in the insect is essential for transmission to the next vertebrate host. The range of insect species that can support the critical development of Plasmodium depends on the individual parasite species, but all five Plasmodium species causing malaria in humans are transmitted exclusively by anopheline mosquitoes. Plasmodium species have remarkable genetic flexibility which lets them adapt to alterations in the environment, giving them the potential to quickly develop resistance to therapeutics such as antimalarials and to change host specificity. In this article, selected topics involving the Plasmodium species that cause malaria in humans are reviewed.
Randomly selected samples from different animal colonies from two laboratory animal houses and from the wild-caught monkeys were tested for the presence of anti-rotavirus antibodies to estimate the rates of infection with group A rotavirus. Antibodies to the common group A rotaviral antigen were detected by a competitive enzyme-linked immunosorbent assay (ELISA) using reagents of WHO ELISA rotavirus detection kit. The results of the study showed that white mice, albino rats, and guinea pigs from long-established breeding colonies and resident house rats and house shrews from the animal house had no serological evidence of rotaviral infection. In contrast, one mousedeer from a colony of 19 animals and most of the rabbits from two separate breeding colonies at the same animal house were serologically positive for the infection. Also a significant number of the same species of monkey kept in captivity were found to acquire the infection. Leaf monkeys had no serological evidence of rotaviral infection. The infection rate in wild cynomolgus monkeys did not seem to be influenced by the different ecological environments of their respective habitats. The rate of infection in adults and juveniles was similar.
Haemosporidians infect a wide diversity of bat genera and species, yet little is known about their transmission cycles or epidemiology. Though several recent studies have focused on the genus Hepatocystis, an Old World parasite primarily infecting bats, monkeys, and squirrels, this group is still understudied with little known about its transmission and molecular ecology. These parasites lack an asexual erythrocytic stage, making them unique from the Plasmodium vertebrate life cycle. In this study, we detected a prevalence of 31% of Hepatocystis in short-nosed fruit bats (Cynopterus brachyotis) in Singapore. Phylogenetic reconstruction with a partial cytochrome b sequence revealed a monophyletic group of Hepatocystis from C. brachyotis in Malaysia, Singapore, and Thailand. There was no relationship with infection and bat age, sex, location, body condition or monsoon season. The absence of this parasite in the five other bat species sampled in Singapore indicates this Hepatocystis species may be host restricted.
Ten years have passed since the publication of a large focus of Plasmodium knowlesi infections in the human population. The discovery was made during a molecular investigation of atypical P. malariae cases in the Kapit Health Division, Sarawak, Malaysian Borneo. Patients were more symptomatic with higher parasite counts than expected in P. malariae infections. The investigation found only P. knowlesi DNA present in patient blood samples. Morphological similarity had allowed P. knowlesi to masquerade as P. malariae during routine diagnostic microscopy for malaria. P. knowlesi, a malaria parasite of macaque monkeys, had entered the human population. The subsequent development of P. knowlesi species-specific PCR assays soon demonstrated that the entry was not confined to the Kapit Division but extended across island and mainland Southeast Asia. Relevant clinical descriptions and guidelines for the treatment and management of patents with P. knowlesi malaria were not available. Nor was it clear whether P. knowlesi had undergone a host switch event into the human population or if infections were zoonotic. The outputs of studies on P. knowlesi malaria during the past 10 years will be summarized, highlighting major findings within the context of pathophysiology, virulence, host switch events, treatment, control and importantly malaria elimination.
Food falsification has direct impact on public health, religious faith, fair-trades and wildlife. For the first time, here we described a multiplex polymerase chain reaction assay for the accurate identification of five meat species forbidden in Islamic foods in a single assay platform. Five pairs of species-specific primers were designed targeting mitochondrial ND5, ATPase 6, and cytochrome b genes to amplify 172, 163, 141, 129 and 108 bp DNA fragments from cat, dog, pig, monkey and rat meats, respectively. All PCR products were identified in gel-images and electrochromatograms obtained from Experion Bioanalyzer. Species-specificity checking against 15 important meat and fish and 5 plant species detected no cross-species amplification. Screening of target species in model and commercial meatballs reflected its application to detect target species in process foods. The assay was tested to detect 0.01-0.02 ng DNA under raw states and 1% suspected meats in meatball formulation.
Ninety-seven specimens of sympatric monkeys and apes from East Malaysia and 115 monkeys and apes from West Africa are examined in order to evaluate the magnitude and nature of the great ape-monkey linear enamel hypoplasia (LEH) 'dichotomy'. This study demonstrates that great apes from both regions have a higher incidence of LEH and repetitive LEH than do gibbons and monkeys. However, the authors find that the dichotomy is not as clear-cut as previous research suggests, since some monkey samples exhibit high LEH frequencies. The authors evaluate the potential influence of great ape-monkey differences in crown height on this dichotomy. They show that canine crown height variation is weakly associated with LEH variation. Differences between monkeys and great apes in their crown formation spans and in their experience of environmental stress may be more likely causes of the dichotomy.
Ancestral sylvatic dengue virus type 1, which was isolated from a monkey in 1972, was isolated from a patient with dengue fever in Malaysia. The virus is neutralized by serum of patients with endemic DENV-1 infection. Rare isolation of this virus suggests a limited spillover infection from an otherwise restricted sylvatic cycle.