Api 20e identification kit

A total of 194 clinical Gram-negative bacterial isolates were recovered from sputum clinical specimens discharged from the microbiology laboratory at Al-Demerdash Hospital, Cairo, Egypt from patients suffering from acute lobar pneumonia according to hospital records during the period from January 2018 to February 2019. Only patients who did not receive previous antimicrobial treatment were included in the study. The isolates were identified using conventional microbiological techniques. Further confirmation of some of the results was done using the API^® 20E identification kit (bioMérieux, Lyon, France). Escherichia coli ATCC^® 25922, E. coli ATCC^® 35218, K. pneumoniae ATCC^® 700603 were used in the quality control of antimicrobial disk diffusion susceptibility tests. The whole study was approved by the Faculty of Pharmacy, Ain Shams University Research Ethics Committee (ENREC-ASU-Nr. 94) where both informed and written consent was obtained from patients or parents of patients after explaining the study purpose.

A total of 165 samples were collected between December 2011 and May 2012 from 18 poultry
markets (mostly street markets and retail shops that sell meat and live birds) distributed
throughout 5 geographical locations in Cairo Governorate. The food samples were collected
from 62 chicken meat parts (20 boneless breasts, 19 cloacae, 10 livers, 8 gizzards and 5
wings), 22 skin pieces from slaughtered birds, 30 raw egg yolks and shells from another 30
eggs. The faecal samples were collected from 21 separate chicken faeces specimens. The
chicken parts and carcass samples were taken from different birds; all samples were
cultured within 2 hours. The faecal samples were obtained from the same 18 poultry
markets.
Salmonella strains were isolated and identified according to standard
methods,¹³ and colonies that
exhibited typical biochemical reactions were further confirmed as
Salmonella using the API 20E identification kit (Biomerieux, Craponne,
France).

The specimens were inoculated onto Nutrient agar plates using a calibrated loop designed to deliver a known volume of 5µl. Inoculated plates were then incubated aerobically at 37°C for 24 hours. After 24 hours, discrete colonies were picked up and gram stained. Further sub-culturing for gram negative bacilli was done on Eosin Methylene Blue agar to obtain a pure culture and biochemical tests were carried out using the Api 20E identification kit in accordance with the manufacturer's manual ((BioMérieux SA, Lyon, France).

The samples were not specifically collected for this research, they were used after the completion of routine laboratory investigations upon the permission of the concerned laboratory's authority. The isolation of bacteria was carried out by conventional bacteriological techniques and identification was confirmed by API 20E identification kit (BioMerieux, France).
^{8 (link)}
As many as 513
K. pneumoniae isolates from various clinical samples and sites (
Figure 1A) were obtained and included in the present study.

Fifty different E. coli isolates were isolated from 50 stool samples of different inpatients carriers, under nonoutbreak conditions, at a hospital in Riyadh, Saudi Arabia, from April 2014 to June 2014. Briefly, fresh stool specimens were aseptically collected and transported to the microbiology laboratory. Stool samples were suspended in sterile phosphate-buffered saline, pH 7. A 100 μL volume was directly inoculated onto blood agar and Eosin Methylene Blue agar (Oxoid Microbiology Products, Hampshire, UK). After 48 h incubation at 37°C, the isolated organisms were identified by conventional procedures and automated identification systems with the API20E identification kit (bioMerieux, Marcy l'Etoile, France). These isolates were preserved in brain heart infusion broth containing 20% glycerol at −70°C.

All the cities, including six cities and three autonomous prefectures in Guizhou province of China, were included in this program. Patients with the following symptoms were selected for the study: Three or more episode of diarrhea within 24 hours, watery or sticky stools, mucus or pus-bloody stools. Also, the following criteria were considered for suspicious cases of non-typhoidal Salmonella infection: fever (temperature >38°C), or with headache, chills, fatigue; nausea, vomiting or abdominal pain. Stool samples from patients with clinical diarrhea were collected to isolate Salmonella from 2013 to 2018. All fecal samples were cultured overnight at 37°C in local hospitals using MacConkey agar plates (Huankai, Guangdong, China). Systematic biochemical methods were used to identify suspected colonies [12 (link)]. All suspicious Salmonella isolates were submitted to the laboratory of Guizhou Provincial Center for Disease Control and Prevention (Guizhou CDC) for further validation and serotyping. In the laboratory, these isolates were recovered by inoculating on nutrient agar plates (Huan Kai, Guangdong, China) and further identified by the API20E identification kit (Biomerieux, France). Identified Salmonella isolates were serotyped by O and H antigen slide agglutination tests (SSI, Denmark) according to the White-Kauffmann-Le Minor scheme [13 ].