Use of biochemical miniaturized galleries, rRNA based lateral flow assay and Real Time PCR for Cronobacter spp. confirmation

Tomás, David; Fan, Mingzhen; Zhu, Sha; Klijn, Adrianne

doi:10.1016/j.fm.2018.05.008

Cited by 2 publications

(2 citation statements)

References 38 publications

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“…In the detection of foodborne pathogens, molecular biological detection technology as a novel identification method has been widely used, which mainly comprises nucleic acid probe techniques, PCR, real-time PCR, and loop-mediated isothermal amplification (LAMP; Wang et al, 2015;Zhang et al, 2015;Tomas et al, 2018;Liang et al, 2019). Among these, the LAMP platform is relatively simple, rapid, and sensitive, which demonstrates greater promise for practical applications in the field (Dhama et al, 2014;Wu et al, 2014;Sarowska et al, 2016;Fu et al, 2020a).…”

Section: Introductionmentioning

confidence: 99%

Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula

Xue

Wang

et al. 2021

Journal of Dairy Science

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Due to the lack of specific genes for rapid detection methods of Cronobacter sakazakii in food samples, whole genome sequence analysis was performed in this investigation using the basic local alignment search tool. Forty-two DNA fragments unique to C. sakazakii were mined, then primers were designed and screened by PCR and loop-mediated isothermal amplification (LAMP). Two primer sets, CS1 and CS31, were found as specific and stable primers, with their corresponding nucleic acid targets the CSK29544_00235 gene and CSK29544_03484 gene, respectively. Furthermore, compared with 3 genes reported previously, these 2 genes were verified as more specific to C. sakazakii among Cronobacter species, by sequence similarity alignment using Cronobacter MLST databases (http: / / pubmlst .org/ cronobacter). The specificity of the LAMP reaction approached 100% by using 48 bacterial strains, which included 22 C. sakazakii strains. Subsequently, LAMP was combined with visual lateral flow dipstick (LFD) based on the above 2 nucleic acid targets, and was demonstrated as a rapid, efficient method with high specificity. Finally, the detection sensitivity of this assay system for pure cultures and artificially contaminated milk was measured as 4.5 × 10 0 cfu/mL and 5.7 × 10 1 cfu/g, respectively. Total time to detection for this assay was within 2 h. Thus, the establishment of this LAMP-LFD method shows great significance and potential for rapid detection of C. sakazakii in powdered infant formula.

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Section: Introductionmentioning

confidence: 99%

Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula

Xue

Wang

et al. 2021

Journal of Dairy Science

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“…bacterial isolates. According toAwong-Taylor et al (2008);Tomas et al (2018), biochemical methods are cheaper, easier to use, give rapid results, and do not require the use of specialized equipment and procedures required for molecular methods. This is especially important when trying to isolate and identify many bacterial isolates in a short period.…”

mentioning

confidence: 99%

Validation of Analytical Method for Aeromonas hydrophila Identification using Analytical Profile Index (API) 20E KIT Method

Artati,

Oman,

Supriyanto

et al. 2024

J. Med. Vet.

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The high demand for fish consumption has an impact on increasing aquaculture productivity and causes the vulnerability of increasing bacterial populations in aquaculture fields, so more rapid handling is needed. The use of phenotypic KIT methods (API 20E) has been applied as a targeted and efficient identification support in targeting better bacterial identification accuracy but often provides unequal results. Method validation is one of the general requirements for the competence of a laboratory evaluation: to provide coherent, interpretable, and accurate results with known uncertainties. The purpose of this study was to validate the API 20E KIT method for the identification of A. hydrophila. The conventional method used as a reference is SNI 7303.1:2015. The validation parameters consisted of determining the limit of detection, sensitivity, and specificity tests, as well as the positive predictive value and negative predictive value. The results showed that the limit of the detection value of the API 20E KIT was at a concentration of 100 CFU/mL with an ID of 99.00%. The sensitivity and specificity values in the positive and negative target samples were 100% with a positive predictive value and a negative predictive value of 0%, respectively. In conclusion, the API 20E KIT method as an alternative test method or rapid test was proven valid for identifying A. hydrophila by the test results using the reference method.

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Use of biochemical miniaturized galleries, rRNA based lateral flow assay and Real Time PCR for Cronobacter spp. confirmation

Cited by 2 publications

References 38 publications

Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula

Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula

Validation of Analytical Method for Aeromonas hydrophila Identification using Analytical Profile Index (API) 20E KIT Method

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