A molecule capturer analysis system for visual determination of avian pathogenic Escherichia coli serotype O78 using a lateral flow assay

Nie, Wenfang; Wang, Juanfang; Xu, Jianguo; Li, Yao; Qiao, Dongqing; Xue, Feng; Tang, Fang

doi:10.1007/s00604-020-4170-6

Cited by 6 publications

(4 citation statements)

References 35 publications

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“…APEC infections are zoonotic in nature and have phylogenic similarities with uropathogenic E. coli (UPEC) that causes urinary tract infections and with neonatal meningitis E. coli that causes neonatal meningitis in humans [16,17]. In addition, these strains can share virulence factors by transmitting virulence genes and plasmids [16]. Furthermore, APEC in meat of healthy birds can be transmitted to humans via the food chain leading to extraintestinal diseases and other diseases [18].…”

Section: Introductionmentioning

confidence: 99%

Virulence Determinants and Multidrug Resistance of Escherichia coli Isolated from Migratory Birds

et al. 2021

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Migratory birds are carriers of multidrug resistant pathogenic Escherichia coli. However, their roles in the dissemination of these resistant pathogens are still being neglected in Bangladesh. The present study was therefore carried out to detect multidrug resistant E. coli. In addition, these isolates were also screened for the presence of avian pathogenic E. coli (APEC)-associated virulence genes. A total of 66 fecal matter samples of migratory birds were screened. E. coli were isolated and identified by culturing and biochemical tests followed by polymerase chain reaction (PCR). APEC-associated virulence genes were detected by PCR. Disk diffusion assays were employed to investigate antibiogram profiles. Bivariate analysis was performed to assess correlations in resistance patterns between antimicrobials and to assess associations between virulence genes of E. coli. Among the 66 samples assessed by PCR, 55 (83.33%) were found positive for E. coli. Of these 55 isolates, the APEC-associated virulence gene fimC was detected in 67.27% of the isolates, which was significantly higher than in the cases of iucD (29.09%) and papC (5.45%) genes. In addition, three isolates were found positive for all three virulence genes, while 23 and 12 isolates were positive for one and two virulence genes respectively. In the bivariate analysis, significant associations were detected between fimC and iucD virulence genes. Using the antibiogram, all E. coli isolates were found to be multidrug resistant (MDR). The isolates exhibited 100% resistance against ampicillin and erythromycin in addition to varying percentages of resistance against streptomycin, tetracycline, ciprofloxacin, and chloramphenicol. Highly positive correlations between tetracycline and ciprofloxacin, chloramphenicol and ciprofloxacin, chloramphenicol and tetracycline were observed by bivariate analysis. To the best of our knowledge, this is the first study that reports APEC-associated virulence genes of MDR E. coli from migratory birds in Bangladesh. Results indicate that migratory birds are reservoirs of MDR E. coli isolates carrying APEC-associated virulence genes, which can seriously contribute to the development of human and animal diseases.

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Section: Introductionmentioning

confidence: 99%

Virulence Determinants and Multidrug Resistance of Escherichia coli Isolated from Migratory Birds

et al. 2021

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“…Along with APEC, uropathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC) are also considered as extra-intestinal pathogenic E. coli (ExPEC). In humans, ExPEC may cause urinary tract infections (UTIs), neonatal meningitis, and septicemia [ 20 , 21 ]. Phylogenic similarities among APEC, UPEC, and NMEC strains—transmissible plasmids, virulence genes, and other genetic characteristics—indicate that APEC are zoonotic in nature [ 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

Molecular Detection of Avian Pathogenic Escherichia coli (APEC) for the First Time in Layer Farms in Bangladesh and Their Antibiotic Resistance Patterns

et al. 2020

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Avian pathogenic Escherichia coli (APEC) causes significant economic losses in poultry industries. Here, we determined for the first time in Bangladesh, the prevalence of APEC-associated virulence genes in E. coli isolated from layer farms and their antibiotic resistance patterns. A total of 99 samples comprising internal organs, feces, and air were collected from 32 layer farms. Isolation was performed by culturing samples on eosin–methylene blue agar plates, while the molecular detection of APEC was performed by PCR, and antibiograms were performed by disk diffusion. Among the samples, 36 were positive for the APEC-associated virulence genes fimC, iucD, and papC. Out of 36 isolates, 7, 18, and 11 were positive, respectively, for three virulence genes (papC, fimC, and iucD), two virulence genes, and a single virulence gene. Although the detection of virulence genes was significantly higher in the internal organs, the air and feces were also positive. The antibiograms revealed that all the isolates (100%) were resistant to ampicillin and tetracycline; 97.2%, to chloramphenicol and erythromycin; 55.5%, to enrofloxacin; 50.0%, to norfloxacin and ciprofloxacin; 19.4%, to streptomycin; 11.1%, to colistin; and 8.33%, to gentamicin. Interestingly, all the isolates were multidrug-resistant (MDR). Spearman’s rank correlation coefficient analysis revealed the strongest significant correlation between norfloxacin and ciprofloxacin resistance. This is the first study in Bangladesh describing the molecular detection of APEC in layer farms. Isolated APEC can now be used for detailed genetic characterization and assessing the impact on public health.

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“…After a series of attempts, the optimum concentration of nanogold-labeled mouse antidigoxigenin antibody was validated to be 10 µg/ml that could be finally selected for better labeling. Thereafter, the LFA strip was established by the method described above (31). In our LFA test strip, the goat anti-biotin antibody and goat anti-mouse IgG antibody were applied with the appropriate concentration for suitable labeling at 1 and 1 mg/ml, respectively.…”

Section: Mechanism and Establishment Of Lateral Flow Assay Strip For Nucleic Acid Detection Of Av Paragallinarummentioning

confidence: 99%

“…In terms of bacteria, this method is also widely applied to detect the nucleic acid of the bacteria. For example, an LFA strip for the detection of avian pathogenic Escherichia coli serotype O78 (APEC O78) is established by Wenfang Nie et al, which can realize the rapid and on-site detection without requiring a sophisticated device and a well-trained operator (31). In terms of Av.…”

Section: Introductionmentioning

confidence: 99%

A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

Huo

et al. 2021

Front. Vet. Sci.

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Avibacterium paragallinarum, the pathogen of infectious coryza, caused a highly contagious respiratory disease that poses a serious threat to chickens. Hence, it is necessary to do diagnostic screening for Av. paragallinarum. Existing technologies have been used for Av. paragallinarum testing, which, however, have some drawbacks such as time consuming and expensive that require well-trained personnel and sophisticated infrastructure, especially when they are limitedly feasible in some places for lack of resources. Nucleic acid hybridization-based lateral flow assay (LFA) is capable of dealing with these drawbacks, which is attributed to the advantages, such low cost, rapid, and simple. However, nucleic acid determination of Av. paragallinarum through LFA method has not been reported so far. In this study, we developed a novel LFA method that employed gold nanoparticle probes to detect amplified Av. paragallinarum dsDNA. Compared with agarose gel electrophoresis, this LFA strip was inexpensive, simple- to- use, and time- saving, which displayed the visual results within 5–8 min. This LFA strip had higher sensitivity that achieved the detection limit of 101 CFU/ml compared with 102 CFU/ml in agarose gel electrophoresis. Besides, great sensitivity was also shown in the LFA strip, and no cross reaction existed for other bacteria. Furthermore, Av. paragallinarum in clinical chickens with infectious coryza were perfectly detected by our established LFA strip. Our study is the first to develop the LFA integrated with amplification and sample preparation techniques for better nucleic acid detection of Av. paragallinarum, which holds great potential for rapid, accurate, and on-site determination methods for early diagnosis of Av. paragallinarum to control further spreading.

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A molecule capturer analysis system for visual determination of avian pathogenic Escherichia coli serotype O78 using a lateral flow assay

Cited by 6 publications

References 35 publications

Virulence Determinants and Multidrug Resistance of Escherichia coli Isolated from Migratory Birds

Virulence Determinants and Multidrug Resistance of Escherichia coli Isolated from Migratory Birds

Molecular Detection of Avian Pathogenic Escherichia coli (APEC) for the First Time in Layer Farms in Bangladesh and Their Antibiotic Resistance Patterns

A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

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