Improved Detection of Mycobacterium tuberculosis in Sudanese Children by use of a Insertion Sequence IS6110 Polymerase Chain Reaction Assay

Abstract

Tuberculosis (TB) is a chronic contagious disease which has a major impact on global public health problem. This study was carried out in Khartoum state during the period from January 2011 to December 2013 to improve detection ofMycobacterium tuberculosisin children with symptoms of tuberculosis infection using different conventional and advanced diagnostic techniques. One hundred ninety seven specimens of gastric lavage and sputum were collected from different hospitals in Khartoum State including ElbolokHospital, Jafar Ibn Owf Hospital, Elasha’ab Teaching Hospital, Soba University Hospital and Academiy Charity Hospital. All childrenparticipating in the study were subjected to mantoux test after obtaining appropriate consent injected by 5 tuberculin units of Tuberculin purified protein derivative and the results were taken after three days.Specimens were decontaminated and inoculated on Lowenstein Jensen mediaaccording to modified Pettrof’s method, Two smears were prepared and stained by Ziehl-Neelsen stain and Auramin fluorescent dye, bacterial DNA was extracted from each specimen by using phenol chloroform method, and then the Polymerase Chain Reaction technique was adopted to detect Insertion Sequence IS6110 gene of M. tuberculosisinthese specimens. This study showed that the positive results for TST, ZN, Auramin, Culture and PCR were 86 (43.7%), 16 (8.1%), 22 (11.2%), 32 (16.2%) and 35(17.8%) respectively. The study concluded that the PCR technique is a most sensitive and specific technique for a fast identification of M. tuberculosisin gastric lavage and sputumfrom children who are unable to expectorate good quality sputum sample or diagnosed as negative using conventional diagnostic methods.