Comparison of Two Different Methods for the Purification of Polymerase Chain Reaction (PCR) Products Used in Direct Sequencing, in an Applied Biosystems 373A DNA Sequencing System

This study was carried out to compare the efficiency of two different purification methods used for removing excess primers and short oligonucleotides from PCR products. Our aim was to sequence PCR product directly. The two different purification methods were micro filter spin unit system (Millipore) and Quick Spin Column (Boehringer Mannheim Biochemica). The results indicated that the purification method using microfilter spin unit system was better and yielded better suquencing pattern compared to quick Spin Column

Avian Influenza (H7N9) Virus Infection in Chinese Tourist in Malaysia, 2014

Of the ≈400 cases of avian influenza (H7N9) diagnosed in China since 2003, the only travel-related cases have been in Hong Kong and Taiwan. Detection of a case in a Chinese tourist in Sabah, Malaysia, highlights the ease with which emerging viral respiratory infections can travel globally

Use of the Reverse Transcriptase Polymerase Chain Reaction for Diagnosis on Dengue Virus Infection Compared to IgM-ELISA

Applicability of the reverse transcriptase polymerase chain reaction (RT-PCR) was evaluated as a routine rapid diagnostic test for dengue virus infection. A total of 160 acute phase sera from patients with clinical diagnosis of dengue fever was examined both by the RT-PCR and IgM-ELISA. Of these, 9 (6%) were positive for both RT-PCR and IgM-ELISA, 61 were positive for IgM-ELISA only and 31 (19%) for RT-PCR only. Both techniques gave negative results in the remaining 59 (37%) specimens. The diagnostic efficiency of IgM-ELISA was statistically better than the RT-PCR even when the specimens were collected on the 3rd or 5th days of the disease. Considering the operational cost in the tests, the acute serum specimens should first be tested by the IgM-ELISA, followed by the RT-PCR on the negative specimens in order to increase the diagnostic efficiency with reasonable cost

The use of casemix system to estimate providers` inpatients’ cost of influenza related sari diseases in Malaysia

Introduction; Influenza is an upper respiratory tract infection caused by the influenza virus. It occurs throughout the whole year in Malaysia with occasional outbreaks. Influenza-like illnesses (ILI) are generally treated as outpatient care while the more severe acute respiratory illness (SARI) is managed in patient care. The Casemix system, used in healthcare professionals' practices, may help in estimating the cost of influenza management. Methods; This cross-sectional study extracts the cost of influenza management from different public hospitals in years 2016, 2017 and 2018. Data used was selected based on ICD-10 codes, and the cost was abstracted from Malaysian Diagnostic Related Groups (MyDRG) software. The secondary data were from two sources, an urban teaching hospital Hospital Cancellor Tuanku Muhriz (HCTM) and Ministry of Health (MoH) inpatient hospitals database. The sample size of the study was 586, while a structured data sheet collating patients’ sociodemographic data and cost of admission, per case, was obtained from the MyDRG software. Microsoft Excel and SPSS software were used in the analysis. Results; Most influenza cases (61.8%) are between the ages of 0 and 10, Malays, and had similar gender proportions. Overall, the influenza treatment cost for inpatient care, was RM71,463,989.16 for 2017, RM78,809,565.60 for 2018 and RM143,743,557.48 for 2019. With the estimated 3.69%, 3.7% and 3.75% of GDP year 2016, 2017 and 2018 for health consumption, the three years trend of influenza consume 0.16% to 0.27% of 2017 and 2018’s GDP. The incremental cost was 40% in year 2018. Conclusions Influenza vaccination, health education and treatment compliance should be scaled-up to minimize the cost of influenza management of the public providers

Genotypes of Japanese Encephalitis Virus Isolated in Three States in Malaysia

Two hundred forty nucleotides from the pre-membrane gene region of 12 Japanese encephalitis virus (JEV) strains isolated from three different regions of Malaysia from 1993 to 1994 were sequenced and compared with each other and with the JEV strains from different geographic areas in Asia. These 12 Malaysian isolates were classified into two genotypes. The four JEV strains isolated from Sarawak in 1994 and the four JEV strains isolated from Sepang, Selangor in 1993 were classified into one genotype that included earlier isolated strains from Malaysia (JE-827 from Sarawak in 1968 and WTP/70/22 from Kuala Lumpur in 1970). The four JEV strains from Ipoh, Perak in 1994 were classified into another genotype that included JEV strains isolated from northern Thailand and Cambodia. In an earlier report, 10 JEV strains from Sabak Bernam, Selangor in 1992 were classified into the largest genotype that included strains isolated in temperate regions such as Japan, China, and Taiwan. The data indicate that at least three genotypes of JEV have been circulating in Malaysia

Air flow optimization for energy efficient blower of biosafety cabinet class II A2

An energy efficient Biosafety Cabinet (BSC) has become a big challenge for manufacturers to develop BSC with the highest level of protection. The objective of research is to increase air flow velocity discharge from centrifugal blower. An aerodynamic duct shape inspired by the shape of Peregrine Falcon's wing during diving flight is added to the end of the centrifugal blower. Investigation of air movement is determined by computational fluid dynamics (CFD) simulation. The results showed that air velocity can be increased by double compared to typical manufactured BSC and no air recirculation. As conclusion, a novel design of aerodynamic duct shape successfully developed and proved that air velocity can be increase naturally with same impeller speed. It can contribute in increasing energy efficiency of the centrifugal blower. It is vital to BSC manufacturer and can be apply to Heating, Air Ventilation and Air Conditioning (HVAC) industries. © Published under licence by IOP Publishing Ltd

Acute Encephalitis Associated with SARS-CoV-2 Confirmed in Cerebrospinal Fluid: First Case in Malaysia

A 69-year-old male with hypertension and atrial fibrillation presented with a 4-day history of fever, cough, and breathlessness, and subsequently developed disorientation and confusion for 1 day. On arrival, his Glasgow Coma Scale was 12/15 (E4, V3, and M5). His body temperature was 37.5°C, blood pressure was 136/60 mm Hg, pulse rate was 72 beats/min , and SpO2 was 98%, with a nasal prong of 3 L/min. There were no symptoms of meningism or longtract signs. On the following day he became increasingly restless and breathless, and required a high-flow-rate nasal cannula to maintain adequate oxygenation. A diagnosis of Coronavirus disease-2019 (COVID-19) was established based on positivity when using an antigen rapid testing kit (SD Biosensor, Inc., Suwon, Korea). However, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was not detected when the polymerase chain reaction (PCR) was applied to the nasopharyngeal swab sample. High-resolution CT of his lungs showed subpleural ground-glass opacities with reticulation on the background of chronic lung changes. Noncontrast brain CT only revealed an old right lenticular infarct. The opening pressure in lumbar puncture was 9.5 cmH2O. His cerebrospinal fluid (CSF) was clear, with a protein level of 1.16 g/L, and the CSF/serum glucose ratio was 0.58. A white blood cell count of 50 cells/mm3 with 100% lymphocytes was seen in the CSF. SARS-CoV-2 was detected in his CSF using PCR with a cycle threshold value of 36.08 and the ORF1ab gene. The results of other CSF analyses were unremarkable

Clinical manifestations of dengue in relation to dengue serotype and genotype in Malaysia: a retrospective observational study

Background: Malaysia experienced an unprecedented dengue outbreak from the year 2014 to 2016 that resulted in an enormous increase in the number of cases and mortality as compared to previous years. The causes that attribute to a dengue outbreak can be multifactorial. Viral factors, such as dengue serotype and genotype, are the components of interest in this study. Although only a small number of studies investigated the association between the serotype of dengue virus and clinical manifestations, none of these studies included analyses on dengue genotypes. The present study aims to investigate dengue serotype and genotype-specific clinical characteristics among dengue fever and severe dengue cases from two Malaysian tertiary hospitals between 2014 and mid-2017. Methodology and principal findings: A total of 120 retrospective dengue serum specimens were subjected to serotyping and genotyping by Taqman Real-Time RT-PCR, sequencing and phylogenetic analysis. Subsequently, the dengue serotype and genotype data were statistically analyzed for 101 of 120 corresponding patients’ clinical manifestations to generate a descriptive relation between the genetic components and clinical outcomes of dengue infected patients. During the study period, predominant dengue serotype and genotype were found to be DENV 1 genotype I. Additionally, non-severe clinical manifestations were commonly observed in patients infected with DENV 1 and DENV 3. Meanwhile, patients with DENV 2 infection showed significant warning signs and developed severe dengue (p = 0.007). Cases infected with DENV 2 were also commonly presented with persistent vomiting (p = 0.010), epigastric pain (p = 0.018), plasma leakage (p = 0.004) and shock (p = 0.038). Moreover, myalgia and arthralgia were highly prevalent among DENV 3 infection (p = 0.015; p = 0.014). The comparison of genotype-specific clinical manifestations showed that DENV 2 Cosmopolitan was significantly common among severe dengue patients. An association was also found between genotype I of DENV 3 and myalgia. In a similar vein, genotype III of DENV 3 was significantly common among patients with arthralgia. Conclusion: The current data contended that different dengue serotype and genotype had caused distinct clinical characteristics in infected patients

Molecular epidemiology of dengue type 2 and 3 viruses in Sarawak and Vietnam

Two methods were carried out to study the molecular epidemiology of Sarawak and Vietnam dengue type 2 and 3 viruses. In the first approach, sequencing of the envelope/non-structural 1 gene junction (E/NS1, 240-bp) was done to infer phylogenetic tree while restriction site-specific polymerase chain reaction was carried out as the second approach. Results from both the methods were compared

Epidemiology Study of Human Metapneumovirus in Malaysia among Paediatric Children below 4 years of age, 2012

We have done a specific study on the molecular epidemiology of Human Metapneumovirus cases which was found in Malaysia among paediatric children below 48 months of age from January till December 2012. These paediatric cases were isolated from the rest of those entire positive below 48 months of age. The total number of the positive cases below 48 months of age is 53 cases out of the general population of 438 positive cases. We performed phylogenetic analysis on these patients and a seasonal prevalence hMPV incidence was observed in the month of November. A high level of sequence identity was observed in the A2 subgroup and no amino acid substitution was found compared to the strains observed in Malaysia and other countries. The pairwise distance among the strains belonging to the predominant subgroup A2was0.0925 suggesting highly homologous with seasonal epidemics. Keyword: Molecular epidemiology, human metapneumovirus, polymerase chain reaction (PCR), paediatric, phylogenetic analysis