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Update on laboratory diagnosis of amoebiasis

Saidin S, Othman N, Noordin R

Eur J Clin Microbiol Infect Dis, 2019 Jan;38(1):15-38.
PMID: 30255429 DOI: 10.1007/s10096-018-3379-3

Amoebiasis, an enteric protozoan disease caused by Entamoeba histolytica, is a public health problem in many developing countries, causing up to 100,000 fatal cases annually. Detection of the pathogenic E. histolytica and its differentiation from the non-pathogenic Entamoeba spp. play a crucial role in the clinical management of patients. Laboratory diagnosis of intestinal amoebiasis in developing countries still relies on labour-intensive and insensitive methods involving staining of stool sample and microscopy. Newer and more sensitive methods include a variety of antigen detection ELISAs and rapid tests; however, their diagnostic sensitivity and specificity seem to vary between studies, and some tests do not distinguish among the Entamoeba species. Molecular detection techniques are highly sensitive and specific and isothermal amplification approaches may be developed into field-applicable tests; however, cost is still a barrier for their use as a routine laboratory test method in most endemic areas. Laboratory diagnosis of extraintestinal amoebiasis faces challenges of lack of definitive detection of current infection and commercially available point-of-care tests. For both types of amoebiasis, there is still a need for highly sensitive and specific tests that are rapid and cost-effective for use in developing countries where the disease is prevalent. In recent years, new molecules of diagnostic value are being discovered and new tests developed. The advances in 'omics' technologies are enabling discoveries of new biomarkers that may help distinguish between different infection stages.
Fulltext In Vitro Testing of Potential Entamoeba histolytica Pyruvate Phosphate Dikinase Inhibitors

Saidin S, Othman N, Noordin R

Am J Trop Med Hyg, 2017 Oct;97(4):1204-1213.
PMID: 28820699 DOI: 10.4269/ajtmh.17-0132

Adverse effects and resistance to metronidazole have motivated the search for new antiamoebic agents against Entamoeba histolytica. Control of amoeba growth may be achieved by inhibiting the function of the glycolytic enzyme and pyruvate phosphate dikinase (PPDK). In this study, we screened 10 compounds using an in vitro PPDK enzyme assay. These compounds were selected from a virtual screening of compounds in the National Cancer Institute database. The antiamoebic activity of the selected compounds was also evaluated by determining minimal inhibitory concentrations (MICs) and IC50 values using the nitro-blue tetrazolium reduction assay. Seven of the 10 compounds showed inhibitory activities against the adenosine triphosphate (ATP)/inorganic phosphate binding site of the ATP-grasp domain. Two compounds, NSC349156 (pancratistatin) and NSC228137 (7-ethoxy-4-[4-methylphenyl] sulfonyl-3-oxido-2, 1, 3-benzoxadiazol-3-ium), exhibited inhibitory effects on the growth of E. histolytica trophozoites with MIC values of 25 and 50 μM, and IC50 values of 14 and 20.7 μM, respectively.
Fulltext Detection of Strongyloides stercoralis infection among cancer patients in a major hospital in Kelantan, Malaysia

Zueter AM, Mohamed Z, Abdullah AD, Mohamad N, Arifin N, Othman N, et al.

Singapore Med J, 2014 Jul;55(7):367-71.
PMID: 25091885

INTRODUCTION: Strongyloidiasis is one of the most commonly neglected but clinically important parasitic infections worldwide, especially among immunocompromised patients. Evidence of infection among immunocompromised patients in Malaysia is, however, lacking. In this study, microscopy, real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISAs) were used to detect Strongyloides stercoralis (S. stercoralis) infection among cancer patients in a Malaysian hospital.

METHODS: A total of 192 stool and serum samples were collected from cancer patients who were receiving chemotherapy with or without steroid treatment at a hospital in northeastern Malaysia. Stool samples were examined for S. stercoralis using parasitological methods and real-time PCR. Serology by ELISA was performed to detect parasite-specific immunoglobulin G (IgG), IgG4 and immunoglobulin E (IgE) antibodies. For comparison, IgG4- and IgG-ELISAs were also performed on the sera of 150 healthy individuals from the same area.

RESULTS: Of the 192 samples examined, 1 (0.5%) sample was positive for S. stercoralis by microscopy, 3 (1.6%) by real-time PCR, 8 (4.2%) by IgG-ELISA, 6 (3.1%) by IgG4-ELISA, and none was positive by IgE-ELISA. In comparison, healthy blood donors had significantly lower prevalence of parasite-specific IgG (2.67%, p < 0.05) and IgG4 (2.67%, p < 0.05) responses.

CONCLUSION: This study showed that laboratory testing may be considered as a diagnostic investigation for S. stercoralis among immunocompromised cancer patients.
Fulltext Analysis of Entamoeba histolytica excretory-secretory antigen and identification of a new potential diagnostic marker

Wong WK, Tan ZN, Othman N, Lim BH, Mohamed Z, Olivos Garcia A, et al.

Clin Vaccine Immunol, 2011 Nov;18(11):1913-7.
PMID: 21918120 DOI: 10.1128/CVI.05356-11

Serodiagnosis of amoebiasis remains the preferred method for diagnosis of amoebic liver abscess (ALA). However, the commercially available kits are problematic in areas of endemicity due to the persistently high background antibody titers. Human serum samples (n = 38) from patients with ALA who live in areas of endemicity were collected from Hospital Universiti Sains Malaysia during the period of 2008 to 2010. Western blots using excretory-secretory antigen (ESA) collected from axenically grown Entamoeba histolytica were probed with the above serum samples. Seven antigenic proteins of ESA with various reactivities were identified, i.e., 152 kDa, 131 kDa, 123 kDa, 110 kDa, 100 kDa, 82 kDa, and 76 kDa. However, only the 152-kDa and 110-kDa proteins showed sensitivities above 80% in the Western blot analysis. All the antigenic proteins showed undetectable cross-reactivity when probed with healthy human serum samples (n = 30) and serum samples from other infections (n = 33). From the matrix-assisted laser desorption ionization-two-stage time of flight (MALDI-TOF/TOF) analysis, the proteins were identified as heavy subunits of E. histolytica lectin and E. histolytica pyruvate phosphate dikinase, respectively. Use of the E. histolytica lectin for diagnosis of ALA has been well reported by researchers and is being used in commercialized kits. However, this is the first report on the potential use of pyruvate phosphate dikinase for diagnosis of ALA; thus, this molecule merits further evaluation on its diagnostic value using a larger panel of serum samples.
Fulltext A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths

Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, et al.

Am J Trop Med Hyg, 2011 Feb;84(2):338-43.
PMID: 21292911 DOI: 10.4269/ajtmh.2011.10-0499

Soil-transmitted helminth infections remain a major public health burden in low- and middle-income countries. The traditional diagnosis by microscopic examination of fecal samples is insensitive and time-consuming. In this study, a pentaplex real-time polymerase chain reaction (PCR) was evaluated for the simultaneous detection of Ancylostoma, Necator americanus, Ascaris lumbricoides, and Strongyloides stercoralis. The results were compared with those obtained by conventional parasitological diagnostic methods. Real-time PCR was positive in 48 of 77 samples (62.3%) and microscopic examination was positive in six samples (7.8%) only (P < 0.05). In conclusion, the real-time PCR assay described in this study provides a specific and sensitive diagnostic tool for the detection of these four helminth species in epidemiological studies and monitoring of treatment programs.
Fulltext Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess

Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, et al.

BMC Infect Dis, 2014 Apr 04;14:182.
PMID: 24708664 DOI: 10.1186/1471-2334-14-182

BACKGROUND: Amoebic liver abscess (ALA) is the most common clinical manifestation of extraintestinal amoebiasis especially in developing countries, causing up to 100 000 fatal cases annually. Accurate and early diagnosis is important to prevent the disease complications, however its diagnosis still poses many challenges due to the limitations of the available detection tools. Pyruvate phosphate dikinase (PPDK), an excretory-secretory protein of E. histolytica, has been reported as a potential diagnostic marker for ALA, hence it may be exploited in the development of a new test for ALA.
METHODS: Recombinant PPDK (rPPDK) was expressed, purified and evaluated by Western blot. In parallel, recombinant galactose-and-N-acetyl-D-galactosamine inhibitable lectin (Gal/GalNAc lectin) was produced and tested similarly. The protein identity was confirmed by analysis using MALDI-TOF/TOF. A lateral flow dipstick (LFD) test using rPPDK was subsequently developed (rPPDK-LFD) and evaluated for serodiagnosis of ALA.
RESULTS: rPPDK was expressed as soluble protein after 4 hours of induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 30°C. Purification using nickel-nitrilotriacetic acid (Ni-NTA) resin yielded 1.5 mg of rPPDK from 1 L of culture with estimated molecular mass of 98 kDa on SDS-PAGE. Western blots using sera from patients with ALA, healthy individuals and other diseases probed with anti-human IgG4-HRP showed the highest sensitivity (93.3%) and specificity (100%); as compared to blots using IgG and IgG1 as secondary antibodies. Moreover, rPPDK showed better specificity when compared to rGal/GalNAc lectin. In the development of the LFD test, the optimum amount of rPPDK was 0.625 μg per dipstick and the optimum working concentration of colloidal gold conjugated anti-human IgG4 was optical density (OD) 5 (1.7 μg of anti-human IgG4). Evaluation of rPPDK-LFD using ALA patients and controls serum samples showed 87% diagnostic sensitivity and 100% specificity.
CONCLUSION: The developed rPPDK-LFD showed good potential for rapid diagnosis of ALA, and merit further multicentre validation using larger number of serum samples.
Infections in post-tsunami victims

Othman N, Ismail IH, Yip R, Zainuddin Z, Kasim SM, Isa R, et al.

Pediatr Infect Dis J, 2007 Oct;26(10):960-1.
PMID: 17901807 DOI: 10.1097/INF.0b013e3181257234

Two tsunami survivors from Banda Acheh, Sumatra, presented with pyrexia of unknown origin and a nonresolving left-sided empyema, respectively. Both children had mixed infections of tuberculosis and melioidosis; Salmonella typhi was also present in the second patient. Mixed infections are common late sequela complications in post-tsunami victims.
Fulltext Head and neck tuberculosis: The great mimic

Mohamad I, Nik Hassan N, Nik Othman NA

Malays Fam Physician, 2016;11(2-3):38.
PMID: 28461859
Fulltext Otitis externa complicated with chloramphenicol ear drops-induced perichondritis

Mohamad I, Johan K, Hashim H, Nik Othman N

Malays Fam Physician, 2014;9(1):28-9.
PMID: 25606295 MyJurnal

Otitis externa is a common condition of the ear. It is manifested as narrowing of the lumen owing to the edematous swelling of the ear canal lining. Perichondritis may occur independently or as a complication of the otitis externa. We report a case of perichondritis after using a topical ear drop. Changing the medication provides immediate resolution of the condition.
Fulltext Predominance and emergence of clones of hospital-acquired methicillin-resistant Staphylococcus aureus in Malaysia

Ghaznavi-Rad E, Nor Shamsudin M, Sekawi Z, Khoon LY, Aziz MN, Hamat RA, et al.

J Clin Microbiol, 2010 Mar;48(3):867-72.
PMID: 20089756 DOI: 10.1128/JCM.01112-09

We define the epidemiology of predominant and sporadic methicillin-resistant Staphylococcus aureus (MRSA) strains in a central teaching and referral hospital in Kuala Lumpur, Malaysia. This is done on the basis of spa sequencing, multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) typing, and virulence gene profiling. During the period of study, the MRSA prevalence was 44.1%, and 389 MRSA strains were included. The prevalence of MRSA was found to be significantly higher in the patients of Indian ethnicity (P < 0.001). The majority (92.5%) of the isolates belonged to ST-239, spa type t037, and possessed the type III or IIIA SCCmec. The arginine catabolic mobile element (ACME) arcA gene was detected in three (1.05%) ST-239 isolates. We report the first identification of ACME arcA gene-positive ST-239. Apart from this predominant clone, six (1.5%) isolates of ST-22, with two related spa types (t032 and t4184) and a singleton (t3213), carrying type IVh SCCmec, were detected for the first time in Asia. A limited number of community-acquired (CA) MRSA strains were also detected. These included ST-188/t189 (2.1%), ST-1/t127 (2.3%), and ST-7/t091 (1%). Panton-Valentin leukocidin (PVL) was detected in all ST-1 and ST-188 strains and in 0.7% of the ST-239 isolates. The majority of the isolates carried agr I, except that ST-1 strains were agr III positive. Virulence genes seg and sei were seen only among ST-22 isolates. In conclusion, current results revealed the predominance of ST-239-SCCmec III/IIIA and the penetration of ST-22 with different virulence gene profiles. The emergence in Malaysia of novel clones of known epidemic and pathogenic potential should be taken seriously.
Social support and work engagement: a study of Malaysian nurses

Othman N, Nasurdin AM

J Nurs Manag, 2013 Nov;21(8):1083-90.
PMID: 23409702 DOI: 10.1111/j.1365-2834.2012.01448.x

This study addressed the question of whether social support (supervisor support and co-worker support) could contribute to the variance in work engagement.
Job characteristics and staying engaged in work of nurses: Empirical evidence from Malaysia

Othman N, Nasurdin AM

Int J Nurs Sci, 2019 Oct 10;6(4):432-438.
PMID: 31728397 DOI: 10.1016/j.ijnss.2019.09.010

Objective: The purpose of this study is to examine the relationship between job characteristics (job autonomy, job feedback, skill variety, task identity, task significance) and work engagement of nurses in Malaysia.
Methods: A survey using self-administered questionnaires was used to collect data from a sample of 856 staff nurses working in eight public hospitals in Malaysia. A shortened nine-item version of the Utrecht Work Engagement Scale(UWES-9) was used to measure work engagement. The UWES-9 comprises three dimensions, which was measured with three items each: vigor, dedication, and absorption. Job characteristics (job autonomy, job feedback, skill variety, task identity, task significance) were measured with the corresponding subscales of the Job Diagnostic Survey. Each subscale consisted of three items. Hypotheses were tested using hierarchical regression analysis.
Results: Findings indicated that all the five demographic variables (age, marital status, education, organizational tenure, job tenure) were unrelated to work engagement. The results further revealed that job autonomy (β = 0.19, P  0.05), however, did not affect work engagement.
Conclusion: Job autonomy, job feedback, task identity, and task significance are important factors in predicting work engagement. The findings of this study highlighted the need to incorporate these core dimensions in nursing management to foster work engagement.
Fulltext Treatment of Wastewater Using Seaweed: A Review

Arumugam N, Chelliapan S, Kamyab H, Thirugnana S, Othman N, Nasri NS

Int J Environ Res Public Health, 2018 12 13;15(12).
PMID: 30551682 DOI: 10.3390/ijerph15122851

Inadequately treated or untreated wastewater greatly contribute to the release of unwanted toxic contaminants into water bodies. Some of these contaminants are persistent and bioaccumulative, becoming a great concern as they are released into the environment. Despite the abundance of wastewater treatment technologies, the adsorption method overall has proven to be an excellent way to treat wastewater from multiple industry sources. Because of its significant benefits, i.e., easy availability, handling, and higher efficiency with a low cost relative to other treatments, adsorption is opted as the best method to be used. However, biosorption using naturally found seaweeds has been proven to have promising results in removing pollutants, such as dyes from textile, paper, and the printing industry, nitrogen, and phosphorous and phenolic compounds, as well as heavy metals from various sources. Due to its ecofriendly nature together with the availability and inexpensiveness of raw materials, biosorption via seaweed has become an alternative to the existing technologies in removing these pollutants from wastewater effectively. In this article, the use of low-cost adsorbent (seaweed) for the removal of pollutants from wastewater has been reviewed. An extensive table summarises the applicability of seaweed in treating wastewater. Literature reported that the majority of research used simulated wastewater and minor attention has been given to biosorption using seaweed in the treatment of real wastewater.
miR-608 regulates apoptosis in human lung adenocarcinoma via regulation of AKT2

Othman N, Nagoor NH

Int J Oncol, 2017 Dec;51(6):1757-1764.
PMID: 29075783 DOI: 10.3892/ijo.2017.4174

Lung cancer remains a major health problem with a low 5-year survival rate of patients. Recent studies have shown that dysregulation of microRNAs (miRNAs) are prevalent in lung cancer and these aberrations play a significant role in the progression of tumour progression. In the present study, bioinformatics analyses was employed to predict potential miR-608 targets, which are associated with signaling pathways involved in cancer. Luciferase reporter assay identified AKT2 as a novel target of miR-608, and suppression of its protein levels was validated through western blot analysis. Zebrafish embryos were microinjected with cells transfected with miR-608 to elucidate the role of miR-608 in vivo, and immunostained with antibodies to detect activated caspase-3. We present the first evidence that miR-608 behaves as a tumour suppressor in A549 and SK-LU-1 cells through the regulation of AKT2, suggesting that selective targeting of AKT2 via miR-608 may be developed as a potential therapeutic strategy for miRNA-based non-small cell lung cancer (NSCLC) therapy.
Overexpression of miR‑361‑5p plays an oncogenic role in human lung adenocarcinoma through the regulation of SMAD2

Othman N, Nagoor NH

Int J Oncol, 2019 01;54(1):306-314.
PMID: 30365047 DOI: 10.3892/ijo.2018.4602

The silencing of Bcl‑xL in the non‑small cell lung cancer (NSCLC) cell line, A549, downregulates miR‑361‑5p expression. This study aimed to determine the biological effects of miR‑361‑5p on NSCLC, and to elucidate the molecular mechanisms through which apoptosis is regulated. MicroRNA (miRNA or miR) functional analyses were performed via transfection of miR‑361‑5p mimics and inhibitors, demonstrating that the inhibition of miR‑361‑5p induced the apoptosis of NSCLC cells. To elucidate the function of miR‑361‑5p in vivo, cells transfected with miR‑361‑5p inhibitors were microinjected into zebrafish embryos, and immunostained using antibodies to detect the active form of caspase‑3. Co-transfection with siBcl‑xL and miR‑361‑5p mimics illustrated the association between Bcl‑xL, miR‑361‑5p and apoptosis; miR‑361‑5p mimics blocked the apoptosis initiated by siBcl‑xL. Luciferase reporter assays identified mothers against decapentaplegic homolog 2 (SMAD2) as a novel target of miR‑361‑5p and the reduction of its protein level was validated by western blot analysis. To confirm the molecular mechanisms through which apoptosis is regulated, gene rescue experiments revealed that the ectopic expression of SMAD2 attenuated the inhibitory effects on apoptosis induced by miR‑361‑5p. In this study, to the best of our knowledge, we provide the first evidence that miR‑361‑5p functions as an oncomiR in A549 and SK‑LU‑1 cells through the regulation of SMAD2, suggesting that miR‑361‑5p may be employed as a potential therapeutic target for the miRNA-based therapy of NSCLC.
Fulltext The role of microRNAs in the regulation of apoptosis in lung cancer and its application in cancer treatment

Othman N, Nagoor NH

Biomed Res Int, 2014;2014:318030.
PMID: 24999473 DOI: 10.1155/2014/318030

Lung cancer remains to be one of the most common and serious types of cancer worldwide. While treatment is available, the survival rate of this cancer is still critically low due to late stage diagnosis and high frequency of drug resistance, thus highlighting the pressing need for a greater understanding of the molecular mechanisms involved in lung carcinogenesis. Studies in the past years have evidenced that microRNAs (miRNAs) are critical players in the regulation of various biological functions, including apoptosis, which is a process frequently evaded in cancer progression. Recently, miRNAs were demonstrated to possess proapoptotic or antiapoptotic abilities through the targeting of oncogenes or tumor suppressor genes. This review examines the involvement of miRNAs in the apoptotic process of lung cancer and will also touch on the promising evidence supporting the role of miRNAs in regulating sensitivity to anticancer treatment.
Fulltext Influence of technology adoption on farmers' well-being: Systematic literature review and bibliometric analysis

Abdul-Majid M, Zahari SA, Othman N, Nadzri S

Heliyon, 2024 Jan 30;10(2):e24316.
PMID: 38312653 DOI: 10.1016/j.heliyon.2024.e24316

This study aimed at determining the effects of technology adoption on farmers' well-being. Specifically, we analysed and extended the current understanding of the topic by focusing on the concepts of technology adoption and well-being. The data were sourced from 30 papers that were selected based on specific criteria from the WoS and Scopus databases. They were acquired through a systematic search and quality appraisal process. The adopted technology can be grouped into sustainable agricultural practices and innovations, digital agriculture and information technology, precision farming and resource management, financial inclusion and agricultural transactions, remote sensing and satellite technology, and agricultural programs and initiatives. Most papers indicated that technology adoption improved farmers' well-being which was basically measured using productivity and income. The measure however lacked farmers' value judgments, such as happiness. Agricultural technology could have a mixed effect on farmers' well-being, depending on the type of technology adopted and the compatibility of farmers with technology in their agricultural practices. This study will assist researchers in developing an appropriate framework and strategy to improve farmers' welfare, which can alleviate poverty. These may further help policymakers identify the best technological innovations that can improve farmers' economic and social welfare, thereby facilitating the development of plans to meet farmers' needs and interests. Finally, the study suggested future research directions for researchers and academics.
Teratocarcinosarcoma of the nasal cavity and ethmoid

Sharma HS, Abdullah JM, Othman NH, Muhamad M

J Laryngol Otol, 1998 Jul;112(7):682-6.
PMID: 9775307

Sinonasal teratocarcinosarcoma is very unusual malignant neoplasm histologically consisting of an epithelial element and one or more mesenchymal components. This is a report of teratocarcinosarcoma, in a 74-year-old male, involving the right nasal cavity and ethmoids with intracranial extension. The tumour was totally resected via the craniofacial approach and the patient was given post-operative chemotherapy. Extensive tumour necrosis, rapid growth and local destruction are the prominent features of this tumour. The clinical presentation, pathological features and clinical course of this rare malignancy are discussed with a review of the literature.
Fulltext The Promise of the Zebrafish Model for Parkinson's Disease: Today's Science and Tomorrow's Treatment

Razali K, Othman N, Mohd Nasir MH, Doolaanea AA, Kumar J, Ibrahim WN, et al.

Front Genet, 2021;12:655550.
PMID: 33936174 DOI: 10.3389/fgene.2021.655550

The second most prevalent neurodegenerative disorder in the elderly is Parkinson's disease (PD). Its etiology is unclear and there are no available disease-modifying medicines. Therefore, more evidence is required concerning its pathogenesis. The use of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is the basis of most animal models of PD. MPTP is metabolized by monoamine oxidase B (MAO B) to MPP + and induces the loss of dopaminergic neurons in the substantia nigra in mammals. Zebrafish have been commonly used in developmental biology as a model organism, but owing to its perfect mix of properties, it is now emerging as a model for human diseases. Zebrafish (Danio rerio) are cheap and easy to sustain, evolve rapidly, breed transparent embryos in large amounts, and are readily manipulated by different methods, particularly genetic ones. Furthermore, zebrafish are vertebrate species and mammalian findings obtained from zebrafish may be more applicable than those derived from genetic models of invertebrates such as Drosophila melanogaster and Caenorhabditis elegans. The resemblance cannot be taken for granted, however. The goal of the present review article is to highlight the promise of zebrafish as a PD animal model. As its aminergic structures, MPTP mode of action, and PINK1 roles mimic those of mammalians, zebrafish seems to be a viable model for studying PD. The roles of zebrafish MAO, however, vary from those of the two types of MAO present in mammals. The benefits unique to zebrafish, such as the ability to perform large-scale genetic or drug screens, should be exploited in future experiments utilizing zebrafish PD models.
Fulltext Circulation of human respiratory syncytial virus strains among hospitalized children with acute lower respiratory infection in malaysia

Etemadi MR, Sekawi Z, Othman N, Lye MS, Moghaddam FY

Evol Bioinform Online, 2013;9:151-61.
PMID: 23641140 DOI: 10.4137/EBO.S10999

Human respiratory syncytial virus (RSV) is a major viral pathogen associated with acute lower respiratory tract infections (ALRTIs) among hospitalized children. In this study, the genetic diversity of the RSV strains was investigated among nasopharyngeal aspirates (NPA) taken from children less than 5 years of age hospitalized with ALRTIs in Hospital Serdang, Malaysia. A total of 165 NPA samples were tested for the presence of RSV and other respiratory viruses from June until December 2009. RSV was found positive in 83 (50%) of the samples using reverse transcription polymerase chain reaction (RT-PCR). Further classification of 67 RSV strains showed that subgroups A and B comprised 11/67 (16.4%) and 56/67 (83.6%) of the strains, respectively. The second hypervariable region at the carboxyl-terminal of the G gene was amplified and sequenced in order to do phylogenetic study. The phylogenetic relationships of the samples were determined separately for subgroups A and B using neighbor joining (NJ), maximum parsimony (MP), and Bayesian inference (BI). Phylogenetic analysis of the 32 sequenced samples showed that all 9 RSV-A strains were clustered within NA1 genotype while the remaining 23 strains of the RSV-B subgroup could be grouped into a clade consisted of strains with 60-nucleotide duplication region. They were further classified into newly discovered BA10 and BA9 genotypes. The present finding suggests the emergence of RSV genotypes of NA1 and BA. This is the first documentation of the phylogenetic relationship and genetic diversity of RSV strains among hospitalized children diagnosed with ALRTI in Serdang, Malaysia.

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