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

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 ۸۵ young children aged ۱۰-۱۵ years. The virus was identified on ۸۵ blood samples using the CER test Human simplex virus Card test; the results revealed that ۱۲ (۱۴.۱%) specimens were related to virus infection, and the sera of patients showed anti-IgM to HSV-۱ antibodies. HSV-۱ 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 ۱۸-۲۴ hours at ۳۷°C. They were then cultivated for ۱۸-۲۴ hours on various selective media at ۳۷°C. The colony morphology, microscopically, and biochemical testing were used to identify Haemophilus influenzae as a first Identification. Out of ۸۵ clinical specimens, ۶۳ (۷۴.۱%) were positive culture, while ۲۲ (۲۵.۹%) had no growth on culture media; out of ۶۳ specimens, only ۲۲ (۳۴.۹%) isolates belonged to Haemophilus influenzae by biochemical tests, while ۴۱ (۶۵.۱%) related to other types of microorganisms. VITEK ۲ was used to validate bacteria isolates from young children suffering from epiglottitis. The findings indicate that ۲۲ (۳۴.۹%) isolates related to Haemophilus influenzae have been confirmed with an excellent ID message confidence level (۹۴ to ۹۹.۸% likelihood percentage). This method is characterized by quick bacterial detection. DNA was taken from all suspected isolates previously identified as Haemophilus influenzae using the vitek۲ 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 ۲۲ (۱۰۰%) samples of Haemophilus influenzae produced ۱۰۱ bp DNA fragments. For isolates previously identified as Haemophilus influenzae, molecular identification of the ompP gene was performed. The results showed that ۱۲ (or ۵۴.۵ percent) of the ۲۲ isolates tested positive for this virulence gene. When compared to an allelic ladder, the presence of (۴۵۹ bp) bands indicated positive results. In addition, the bexA gene was molecularly detected in ۲۲ Haemophilus influenzae isolates, showing that only ۸ (۳۶.۳ percent) of the isolates had this gene. When compared to an allelic ladder, the presence of a (۳۴۳ bp) band indicated positive results for bexA gene pathogenicity; in conclusion, HSV (۱) and Hib were considered almost causative agents of epiglottitis in young children.

Keywords:

Haemophilus influenzae , viral infection , Bacterial infection , HSV-۱ , epiglottitis