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  1. Optical test strip for trace Hg(II) based on doped sol-gel film
    Yusof NA, Kadir WA
    Spectrochim Acta A Mol Biomol Spectrosc, 2009 Feb;72(1):32-5.
    PMID: 19010723 DOI: 10.1016/j.saa.2008.07.019
    Optical test strip based on the use of Br-PADAP as a sensitive reagent immobilised into sol-gel thin film for detection of Hg(II) in aqueous solution had been thoroughly carried out. It has a square-sensing zone (1.0 cm x 1.0 cm) containing the sensitive reagent necessary to produce response to trace level of mercury. This method offer sensitivity and simplicity in detecting Hg(II) as no prior treatment or extraction is required. A linear response was attained in the Hg(II) concentration in the range of 0.5-2.5 ppm with calculated limit of detection of 6.63 ppb. This method also showed a reproducible result with relative standard deviation (R.S.D.) of 2.15% and response time of approximately 5 min. Interference studies showed that Al(III), Co(II) and Ni(II) significantly interfered during the determination. The developed sensor has been validated against Atomic Absorption Spectroscopy method and proven comparable.
    Matched MeSH terms: Reagent Strips/chemistry*
  2. 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.

    Matched MeSH terms: Reagent Strips/chemistry*
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