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  1. Mohd Ali MR, Lih Huey L, Foo PC, Goay YX, Ismail AS, Mustaffa KMF, et al.
    Biomed Res Int, 2019;2019:9451791.
    PMID: 31355287 DOI: 10.1155/2019/9451791
    Melioidosis and leptospirosis, caused by two different bacteria, Burkholderia pseudomallei and Leptospira spp., are potentially fatal infections that share a very similar spectrum of clinical features and cause significant mortality and morbidity in humans and livestock. Early detection is important for better clinical consequences. To our knowledge, there is no diagnostic tool available to simultaneously detect and differentiate melioidosis and leptospirosis in humans and animals. In this study, we described a duplex TaqMan probe-based qPCR for the detection of B. pseudomallei and Leptospira spp. DNA. The performance of the assay was evaluated on 20 B. pseudomallei isolates, 23 Leptospira strains, and 39 other microorganisms, as well as two sets of serially diluted reference strains. The duplex qPCR assay was able to detect 0.02 pg (~ 4 copies) Leptospira spp. DNA and 0.2 pg (~ 25.6 copies) B. pseudomallei DNA. No undesired amplification was observed in other microorganisms. In conclusion, the duplex qPCR assay was sensitive and specific for the detection of B. pseudomallei & Leptospira spp. DNA and is suitable for further analytical and clinical evaluation.
  2. Nurul Najian AB, Foo PC, Ismail N, Kim-Fatt L, Yean CY
    Mol Cell Probes, 2019 04;44:63-68.
    PMID: 30876924 DOI: 10.1016/j.mcp.2019.03.001
    This study highlighted the performance of the developed integrated loop-mediated isothermal amplification (LAMP) coupled with a colorimetric DNA-based magnetogenosensor. The biosensor operates through a DNA hybridization system in which a specific designed probe captures the target LAMP amplicons. We demonstrated the magnetogenosensor assay by detecting pathogenic Leptospira, which causes leptospirosis. The color change of the assay from brown to blue indicated a positive result, whereas a negative result was indicated by the assay maintaining its brown color. The DNA biosensor was able to detect DNA at a concentration as low as 200 fg/μl, which is equivalent to 80 genomes/reaction. The specificity of the biosensor assay was 100% when it was evaluated with 172 bacterial strains. An integrated LAMP and probe-specific magnetogenosensor was successfully developed, promising simple and rapid visual detection in clinical diagnostics and service as a point-of-care device.
  3. Engku Nur Syafirah EAR, Nurul Najian AB, Foo PC, Mohd Ali MR, Mohamed M, Yean CY
    Acta Trop, 2018 Jun;182:223-231.
    PMID: 29545156 DOI: 10.1016/j.actatropica.2018.03.004
    Cholera, caused by Vibrio cholerae is a foodborne disease that frequently reported in food and water related outbreak. Rapid diagnosis of cholera infection is important to avoid potential spread of disease. Among available diagnostic platforms, loop-mediated isothermal amplification (LAMP) is regarded as a potential diagnostic tool due to its rapidity, high sensitivity and specificity and independent of sophisticated thermalcycler. However, the current LAMP often requires multiple pipetting steps, hence is susceptible to cross contamination. Besides, the strict requirement of cold-chain during transportation and storage make its application in low resource settings to be inconvenient. To overcome these problems, the present study is aimed to develop an ambient-temperature-stable and ready-to-use LAMP assay for the detection of toxigenic Vibrio cholerae in low resource settings. A set of specific LAMP primers were designed and tested against 155 V. cholerae and non-V. cholerae strains. Analytical specifity showed that the developed LAMP assay detected 100% of pathogenic V. cholerae and did not amplified other tested bacterial strains. Upon testing against stool samples spiked with toxigenic V. cholerae outbreak isolates, the LAMP assay detected all of the spiked samples (n = 76/76, 100%), in contrast to the conventional PCR which amplified 77.6% (n = 59/76) of the tested specimens. In term of sensitivity, the LAMP assay was 100-fold more sensitive as compared to the conventional PCR method, with LOD of 10 fg per μL and 10 CFU per mL. Following lyophilisation with addition of lyoprotectants, the dry-reagent LAMP mix has an estimated shelf-life of 90.75 days at room temperature.
  4. Wong WK, Mohd-Nor N, Noordin R, Foo PC, Mohamed Z, Haq JA, et al.
    Parasitol Res, 2019 Sep;118(9):2635-2642.
    PMID: 31363922 DOI: 10.1007/s00436-019-06406-7
    The geographical distribution of tuberculosis (TB) overlaps with various parasitic infections. Uncovering the characteristics of coinfecting parasites that potentially affect the host susceptibility to TB is pertinent as it may provide input to current TB therapeutic and prophylactic measures. The present study was aimed at examining the types of parasitic infections in TB patients and healthy TB contacts (HC) in Orang Asli, Malaysian aborigines, who dwelled in the co-endemic areas. Stool and serum samples were collected from Orang Asli who fulfilled the selection criteria and provided written informed consents. Selected parasitic infections in the two study groups were determined by stool examination and commercial serum antibody immunoassays. The prevalence of parasitic infections in TB and HC participants were 100% (n = 82) and 94.6% (n = 55) respectively. The parasitic infections comprised toxocariasis, trichuriasis, amoebiasis, toxoplasmosis, hookworm infection, ascariasis, strongyloidiasis, and brugian filariasis, in decreasing order of prevalence. Overall, helminth or protozoa infection did not show any significant association with the study groups. However, when the species of the parasite was considered, individuals exposed to trichuriasis and toxoplasmosis showed significant odds reduction (odds ratio (OR) 0.338; 95% confidence interval (CI) 0.166, 0.688) and odds increment (OR 2.193; 95% CI 1.051, 4.576) to have active pulmonary TB, respectively. In conclusion, trichuriasis and toxoplasmosis may have distinct negative and positive associations respectively with the increase of host susceptibility to TB.
  5. Foo PC, Nurul Najian AB, Muhamad NA, Ahamad M, Mohamed M, Yean Yean C, et al.
    BMC Biotechnol, 2020 Jun 22;20(1):34.
    PMID: 32571286 DOI: 10.1186/s12896-020-00629-8
    BACKGROUND: This study reports the analytical sensitivity and specificity of a Loop-mediated isothermal amplification (LAMP) and compares its amplification performance with conventional PCR, nested PCR (nPCR) and real-time PCR (qPCR). All the assays demonstrated in this study were developed based on Serine-rich Entamoeba histolytica protein (SREHP) gene as study model.

    RESULTS: A set of SREHP gene specific LAMP primers were designed for the specific detection of Entamoeba histolytica. This set of primers recorded 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. These primers were later modified for conventional PCR, nPCR and qPCR applications. Besides, 3 different post-LAMP analyses including agarose gel electrophoresis, nucleic acid lateral flow immunoassay and calcein-manganese dye techniques were used to compare their limit of detection (LoD). One E. histolytica trophozoite was recorded as the LoD for all the 3 post-LAMP analysis methods when tested with E. histolytica DNA extracted from spiked stool samples. In contrast, none of the PCR method outperformed LAMP as both qPCR and nPCR recorded LoD of 100 trophozoites while the LoD of conventional PCR was 1000 trophozoites.

    CONCLUSIONS: The analytical sensitivity comparison among the conventional PCR, nPCR, qPCR and LAMP reveals that the LAMP outperformed the others in terms of LoD and amplification time. Hence, LAMP is a relevant alternative DNA-based amplification platform for sensitive and specific detection of pathogens.

  6. Foo PC, Chan YY, See Too WC, Tan ZN, Wong WK, Lalitha P, et al.
    J Med Microbiol, 2012 Sep;61(Pt 9):1219-1225.
    PMID: 22556327 DOI: 10.1099/jmm.0.044552-0
    Entamoeba histolytica is the only Entamoeba species that causes amoebiasis in humans. Approximately 50 million people are infected, with 100, 000 deaths annually in endemic countries. Molecular diagnosis of Entamoeba histolytica is important to differentiate it from the morphologically identical Entamoeba dispar to avoid unnecessary medication. Conventional molecular diagnostic tests require trained personnel, cold-chain transportation and/or are storage-dependent, which make them user-unfriendly. The aim of this study was to develop a thermostabilized, one-step, nested, tetraplex PCR assay for the detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba species in cold-chain-free and ready-to-use form. The PCR test was designed based on the Entamoeba small subunit rRNA (SSU-rRNA) gene, which detects the presence of any Entamoeba species, and simultaneously can be used to differentiate Entamoeba histolytica from Entamoeba dispar. In addition, a pair of primers was designed to serve as an internal amplification control to help identify inhibitors in the samples. All PCR reagents together with the designed primers were thermostabilized by lyophilization and were stable at 24 °C for at least 6 months. The limit of detection of the tetraplex PCR was found to be 39 pg DNA or 1000 cells for Entamoeba histolytica and 78 pg DNA or 1000 cells for Entamoeba dispar, and the specificity was 100 %. In conclusion, this cold-chain-free, thermostabilized, one-step, nested, multiplex PCR assay was found to be efficacious in differentiating Entamoeba histolytica from other non-pathogenic Entamoeba species.
  7. Wong WK, Foo PC, Roze MN, Pim CD, Subramaniam P, Lim BH
    Can J Infect Dis Med Microbiol, 2016;2016:1326085.
    PMID: 27366156 DOI: 10.1155/2016/1326085
    Background. Orang Asli (aborigine) children are susceptible to soil-transmitted helminth (STH) infections due to their lifestyle and substandard sanitation system. Objectives. This study aimed to examine the helminthic and nutritional status of Orang Asli school children in Sekolah Kebangsaan Pos Legap, a remote primary school at Kuala Kangsar District in the state of Perak, Malaysia. In addition, the sensitivities of four STH stool examination techniques were also compared. Methods. Demography and anthropometry data were collected by one-to-one interview session. Collected stools were examined with four microscopy techniques, namely, direct wet mount, formalin ether concentration (FEC), Kato-Katz (KK), and Parasep™. Results. Anthropometry analysis showed that 78% (26/33) of children in SK Pos Legap were malnourished and 33% (11/33) of them were stunted. Stool examinations revealed almost all children (97%) were infected by either one of the three commonest STHs. FEC was the most sensitive method in detection of the three helminth species. Conclusion. This study revealed that STH infections and nutritional status still remain a health concern among the Orang Asli children. These communal problems could be effectively controlled by regular monitoring of STH infection loads, administration of effective antihelminthic drug regimen, and also implementation of effective school nutritional programs.
  8. Foo PC, Chan YY, Mohamed M, Wong WK, Nurul Najian AB, Lim BH
    Anal Chim Acta, 2017 May 08;966:71-80.
    PMID: 28372729 DOI: 10.1016/j.aca.2017.02.019
    This study highlighted the development of a four target nitrocellulose-based nucleic acid lateral flow immunoassay biosensor in a dry-reagent strip format for interpretation of double-labelled double-stranded amplicons from thermostabilised triplex loop-mediated isothermal amplification assay. The DNA biosensor contained two test lines which captured biotin and texas red labelled amplicons; a LAMP internal amplification control line that captured digoxigenin labelled amplicon; and a chromatography control line that validated the functionality of the conjugated gold nanoparticles and membrane. The red lines on detection pad were generated when the gold nanoparticles conjugated antibody bound to the fluorescein labelled amplicons, and the capture agents bound to their specific hapten on the other 5' end of the double-stranded amplicon. The applicability of this DNA biosensor was demonstrated using amoebiasis-causing Entamoeba histolytica simultaneously with the non-pathogenic but morphologically identical Entamoeba dispar and Entamoeba moshkovskii. The biosensor detection limit was 10 E. histolytica trophozoites, and revealed 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. Heat stability test showed that the biosensor was stable for at least 181 days at ambient temperature. This ready-to-use and cold-chain-free biosensor facilitated the post-LAMP analysis based on visualisation of lines on strip instead of observation of amplicon patterns in agarose gel.
  9. Wong WK, Foo PC, Olivos-Garcia A, Noordin R, Mohamed Z, Othman N, et al.
    Acta Trop, 2017 Aug;172:208-212.
    PMID: 28506795 DOI: 10.1016/j.actatropica.2017.05.017
    Crude soluble antigen (CSA) produced from Entamoeba histolytica trophozoite is conventionally used for serodiagnosis of invasive amoebiasis. However, high background seropositivities by CSA-assay in endemic areas complicate the interpretation of positive result in clinical settings. Instead, incorporating a second assay which indicates active or recent infection into the routine amoebic serology could possibly complement the limitations of CSA-assay. Hence, the present study aimed to evaluate the diagnostic efficacies of indirect ELISAs using CSA and excretory-secretory antigen (ESA) for serodiagnosis of amoebic liver abscess (ALA). Reference standard for diagnosis of ALA at Hospital Universiti Sains Malaysia is based on clinical presentation, radiological imaging and positive indirect haemagglutination assay (titer ≥256). Five groups of human serum samples collected from the hospital included Group I - ALA diagnosed by the reference standard and pus aspirate analysis using real-time PCR (n=10), Group II - ALA diagnosed by the reference standard only (n=41), Group III - healthy control (n=45), Group IV - other diseases control (n=51) and Group V - other infectious diseases control (n=31). For serodiagnosis of ALA serum samples (Group I and II), CSA-ELISA showed sensitivities of 100% for both groups, while ESA-ELISA showed sensitivities of 100% and 88%, respectively. For serodiagnosis of non-ALA serum samples (Group III, IV and V), CSA-ELISA showed specificities of 91%, 75% and 100%, respectively; while ESA-ELISA showed specificities of 96%, 98% and 100%, respectively. Indirect ELISAs using CSA and ESA have shown distinct strength for serodiagnosis of ALA, in terms of sensitivity and specificity, respectively. In conclusion, parallel analysis by both assays improved the overall efficacies of amoebic serology as compared to either single assay.
  10. Ahmad NI, Yean Yean C, Foo PC, Mohamad Safiee AW, Hassan SA
    J Infect Public Health, 2020 Oct;13(10):1508-1512.
    PMID: 32653480 DOI: 10.1016/j.jiph.2020.06.018
    BACKGROUND: Panton-Valentine Leukocidin (PVL), is one of the virulence gene expressed by Methicillin Resistant Staphylococcus aureus (MRSA) and is known to be associated with severe form of community acquired MRSA infection. The aim of this study is to investigate its prevalence in our setting and patient's clinical outcome.

    METHODS: A cross sectional study involve retrospective record review were done involving 90 MRSA positive isolates between November 2016 and October 2017. Multiplex PCR was performed to detect femA, mecA and PVL genes. Clinical presentation and outcomes of patients were reviewed and presented as descriptive analysis.

    RESULTS: All of the 90 MRSA isolates included in this study were positive for femA and mecA genes following PCR. PVL gene was detected in 20% (n = 18) of the isolates of which 61.1% (n = 11) were community acquired infections and 38.8% (n = 7) were hospital acquired. Case distribution from community acquired infections include patients with skin and soft tissue infections (33.3%, n = 6), infected diabetic foot ulcers (16.7%, n = 3), and one patient each (5.5%, n = 1) for community acquired pneumonia and meningitis. Half of the PVL positive MRSA cases (50%, n = 9) were having sepsis and four of them succumbed to death due to severe infection.

    CONCLUSION: This study shows a high prevalence of PVL positive MRSA infection in our population. Skin and soft tissue infections accounting for the major sources. In addition, the presence of the PVL gene is associated with increased risk for developing sepsis.

  11. Foo PC, Lee ZH, Chin CK, Subramaniam S, Chew BL
    Trop Life Sci Res, 2018 Jul;29(2):119-129.
    PMID: 30112145 MyJurnal DOI: 10.21315/tlsr2018.29.2.9
    Solanum melongena L. commonly known as the eggplant or brinjal comes from the family of Solanaceae, sharing the same ancestor with the tomato and potato. It is an economically important crop worldwide, being well studied for its medicinal properties, nutritional values and its role as an alternative model plant. The eggplant fruit has been previously used for treatments of various diseases such as bronchitis, asthma, arthritis and diabetes as well as its nutritive properties that are beneficial to the human diet. Plant transformation studies on the eggplant have been widely done for the production of transgenic eggplants harbouring genes that are beneficial for optimal plant growth and fruit production. Shoot induction is an essential step required for the successful regeneration of transformed plant tissues and therefore is an essential pre-requisite in Agrobacterium-mediated transformation. The local eggplant cv. Bulat Putih is a local cultivar of eggplant in Malaysia with white and round fruits making it a potential model plant colour pigment accumulation studies in fruit crops. The current work aims to investigate the shoot induction potential of 6-benzylaminopurine (BAP) and Kinetin from cotyledon explants of eggplant cv. Bulat Putih. Results indicated that both BAP and Kinetin were able to induce the regeneration of callus from cotyledon explants. On the other hand, Kinetin at the concentration of 2.0 mg/L successfully induced shoots at the value of 1.50 ± 0.22 shoots per explant, whereas BAP alone did not trigger any formation of shoots. This study indicated that kinetin alone is sufficient to induce shoots in eggplant cv. Bulat Putih without the presence of BAP.
  12. Nurhafiza NNBA, Siti Asma H, Azian H, Foo PC, Yasmin KMI, Chan YY
    Singapore Med J, 2020 12 02.
    PMID: 33264563 DOI: 10.11622/smedj.2020166
    INTRODUCTION: This study determined the distribution of sasX, qacA/B and mupA genes from methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical samples and nasal swab samples of the same patients and analysed their genetic relatedness.

    METHODS: Polymerase chain reaction (PCR) was used to detect the presence of sasX, qacA/B and mupA genes from 47 paired MRSA isolates. A paired isolate was defined as one nasal swab (colonising) isolate and clinical isolate that caused infection in the same patient. 22 selected paired isolates were subjected to multilocus sequence typing (MLST). The genetic relatedness among the isolates and association between the putative genes with epidemic sequence types (STs) were investigated.

    RESULTS: 7 (14.9%, n = 14) paired isolates were positive for the sasX gene. qacA/B genes were positive in 7.4% (n = 7) of the isolates, from three paired isolates and one clinical isolate whose paired colonising isolate was negative. The paired sample of three patients were positive for both genes. The mupA gene was not detected in all the isolates. MLST revealed two epidemic STs, ST22 and ST239, and a novel ST4649. sasX and qacA/B genes were found in ST239 in 29.5% (n = 13) and 13.6% (n = 6) of cases, respectively. Gene co-existence occurred in 13.6% (n = 6) of MRSA ST239 and 2.3% (n = 1) of MRSA ST4649.

    CONCLUSION: sasX and qacA/B genes were present in the MRSA isolates, while the mupA gene was undetected. ST22 and ST239 were the major MRSA clones. The circulating MRSA genotypes conferred different virulence and resistance determinants in our healthcare settings.

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