Detection of Main Causative Agents among Young Children Suffering from Epiglottitis in Hilla City, Iraq.

Abstract

Epiglottitis is a rapidly progressive epiglottis infection leading to upper airway edema. This study aimed to detect the main causative agent, viral infection, by immunofluorescence antibody technique and PCR technique and bacterial infection detection by specific gene among young children suffering from epiglottitis. This study included 85 young children aged 10-15 years. The virus was identified on 85 blood samples using the CER test Human simplex virus Card test; the results revealed that 12 (14.1%) specimens were related to virus infection, and the sera of patients showed anti-IgM to HSV-1 antibodies. HSV-1 was detected in blood samples by qPCR technique. Eighty-five saliva samples were collected from young children suffering from epiglottitis. The samples were cultured for 18-24 hours at 37°C. They were then cultivated for 18-24 hours on various selective media at 37°C. The colony morphology, microscopically, and biochemical testing were used to identify Haemophilus influenzae as a first Identification. Out of 85 clinical specimens, 63 (74.1%) were positive culture, while 22 (25.9%) had no growth on culture media; out of 63 specimens, only 22 (34.9%) isolates belonged to Haemophilus influenzae by biochemical tests, while 41 (65.1%) related to other types of microorganisms. VITEK 2 was used to validate bacteria isolates from young children suffering from epiglottitis. The findings indicate that 22 (34.9%) isolates related to Haemophilus influenzae have been confirmed with an excellent ID message confidence level (94 to 99.8% likelihood percentage). This method is characterized by quick bacterial detection. DNA was taken from all suspected isolates previously identified as Haemophilus influenzae using the vitek2 technology, and traditional PCR was used to amplify specific hel gene for Haemophilus influenzae primers utilizing these DNA samples. After that, when compared to an allelic ladder, gel electrophoresis revealed that all 22 (100%) samples of Haemophilus influenzae produced 101 bp DNA fragments. For isolates previously identified as Haemophilus influenzae, molecular identification of the ompP gene was performed. The results showed that 12 (or 54.5 percent) of the 22 isolates tested positive for this virulence gene. When compared to an allelic ladder, the presence of (459 bp) bands indicated positive results. In addition, the bexA gene was molecularly detected in 22 Haemophilus influenzae isolates, showing that only 8 (36.3 percent) of the isolates had this gene. When compared to an allelic ladder, the presence of a (343 bp) band indicated positive results for bexA gene pathogenicity; in conclusion, HSV (1) and Hib were considered almost causative agents of epiglottitis in young children.