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Abstrak :
ABSTRACT
Hepatitis B virus (HBV) infects hepatocytes and causes acute and chronic hepatitis that can lead to cirrhosis and hepatocellular carcinoma (HCC) in both animals and humans. Early detection of HBV infection assists in monitoring the patient's response to anti-HBV therapy, blood donation screening, and disease management, control and eradication. This research focused on development of LAMP assay combined with lateral flow dipstick (LFD), gold nanoparticle (AuNPs) and real-time turbidimetry for screening of the hepatitis B virus. Analytical sensitivity, analytical specificity, diagnostic sensitivity, diagnostic specificity, accuracy and predictive value of each technique were determined and compared to conventional PCR and real-time PCR (gold standard method). The analytical sensitivity of LAMP-LFD and LAMP-AuNPs was 1.24x101 copies /mL, LAMP-real-time turbidimetry was 1.24x102 copies/mL, while that of conventional PCR was 1.24x104 copies/mL. Examination of the analytical specificity of all LAMP-based combinations and conventional PCR showed no cross-reactivity with HCV or human plasma. Upon exploration of one hundred unknown samples, in comparison to real-time PCR, the diagnostic sensitivity and specificity of LAMP-based assays were 100% and 90%, respectively. The accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the LAMP-based assays were 98%, 97.56%, and 100%, respectively. While that of conventional PCR were 60%, 100%, 68%, 100% and 38% of diagnostic sensitivity, diagnostic specificity, accuracy, PPV and NPV, respectively. LAMP-based assays need to be simplified in terms of achieving single-step diagnosis using one master mix solution that is suitable for a point-of-care diagnostic test.

Abstrak :
ABSTRAK
Development of the Rapid Test Kit for the Identification of Campylobacter spp. Based on Loop-mediated Isothermal Amplification (LAMP) in Combination with a Lateral Flow Dipstick (LFD) and Gold Nano-DNA Probe (AuNPs) Dueantem Thongphueak, Kosum Chansiri Center of Excellence in Biosensors, Srinakharinwirot University, Bangkok 10110, Thailand. Thayat Sriyapai Faculty of Environmental Culture and Ecotourism, Srinakharinwirot University, Bangkok 10110, Thailand. Supatra Areekit, Somchai Santiwatanakul, Piyada Wangroongsarb ABSTRAK
The detection of Campylobacter spp. in meat products was developed by using loop-mediated isothermal amplification (LAMP) combined with DNA-based bioassay methods, including a lateral-flow dipstick (LFD) and gold nano-DNA probe (AuNPs) assay. The LAMP primers were designed from the conserved nucleotide regions of Campylobacter spp. The analytical sensitivity of the LAMP-LFD and LAMP-AuNPs analysis was 360 fg/μl. The analytical specificity of LAMP-based assays showed no cross-reactions to Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli, Bacillus cereus, Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacter aerogenes, Serratia marcescens, Vibrio parahaemolyticus, Vibrio cholerae, Klebsiella oxytoca and Citrobacter diversus.The sensitivity, specificity and accuracy of both LAMP-LFD and LAMP-AuNPs for the detection of pre-enrichment cultures from raw chicken meat samples were 100%, 95% and 96.67%, respectively. Since the processing time of LAMP-based assays is 60-90 minutes, it is applicable as a point-of-care screening test for food safety and as a process control of Campylobacter spp. contamination.