Main Article Content
This is a case study of the incidence of diphtheria in National Ear Care Centre Kaduna. Samples included all patients presented with diphtheria symptoms referred to the hospital from October to December; 2018. A total of 8 patients were referred and several samples taken for laboratory investigations. 8 throat swabs were collected for organism isolation by inoculation on some certain agars, incubation, secondary gram and microscopy; colonies appeared to be biconcave, large and whitish. Microscopy revealed characteristic Chinese lettered, pleomorphic bacillus which are features of Corynebacterium diphtheriae. Blood samples were used for urea and creatinine estimation which yielded abnormal high values in all except the creatinine of one of the samples indicating kidney impairment which could be attributed to presence of the tox gene. To further confirm the incidence of the disease; one throat swab was saved at 4°C for molecular confirmation and characterisation of the gene. It was then transported to Molecular laboratory and the DNA extracted using the phenol/chloroform extraction method. Both the A and B subunits of the tox gene were amplified from the DNA extract at 248bp and 297bp respectively. Sanger’s Dye Terminator Cycle Sequencing (DTCS) of both the subunits of the tox gene was carried out and the sequences were obtained and converted to the fasta format using a Finch TV for bioinformatics analyses using the NCBI blast tool. The A and B subunits showed a percentage identity of 99.59% and 95.57% to Corynebacterium diphtheriae mitis strain NCTC 3529 genome assembly, chromosome: 1 respectively. The A subunit also showed 99.59% identity to Corynebacterium diphtheriae strain BQ11 chromosome; complete genome while the B subunit also showed 95.57% identity to Corynebacterium diphtheriae mitis strain NCTC 7838 genome assembly chromosome: 1. A mutation was also discovered in the tox genes as indicated by the E-value of 3e-84 and 2e-121 for A and B subunits respectively which are values that are less than 1. This is also confirmed by 5 gaps noticed from the blast results (4 gaps from A and 1 from B) indicating the presence of indels. In conclusion; there are confirmed incidences and sporadic cases of diphtheria in Kaduna State and the organism is said to have mutated.
Klebs E. III. Sitzung: Ueber Diphtherie (Third session: On diphtheria), Verhandlungen des Congresses für innere Medicin. Zweiter Congress gehalten zu Wiesbaden, 18.-23. (Proceedings of the congress on internal medicine. Second congress held at Wiesbaden. 1883;2:139-154.
Cerdeño-Tárraga AM, Efstratiou A, Dover LG, Holden MT, Pallen M, Bentley SD, Besra GS, Churcher C, James KD, De Zoysa A, Chillingworth T, Cronin A, Dowd L, Feltwell T, Hamlin N, Holroyd S, Jagels K, Moule S, Quail MA, Rabbinowitsch E, Rutherford KM, Thomson NR, Unwin L, Whitehead S, Barrell BG, Parkhill J. The complete genome sequence and analysis of Corynebacterium diphtheriae. Nucleic Acids Research. 2003;22:6516–23.
Sangal V, Tucker NP, Burkovski A, Hoskisson PA. The draft genome sequence of Corynebacterium diphtheriae bv. Mitis NCTC 3529 reveals significant diversity between the primary disease-causing biovars. Journal of Bacteriology. 2012;194(12):3269.
Chakraborty SM, Choudghuri AB. Isolation of toxigenic strains of C. diphtheriae from skin lesions of children in a rural West Bengal community. Indian Journal of Medical Research. 1969;57:1651-8.
Atkinson W, Hamborsky J, McIntyre L, Wolfe S. Diphtheria. Epidemiology And Prevention of Vaccine-Preventable Diseases (10 ed.). Public Health Foundation. 2007;59–70.
Behrman RE, Kliegman RM, Jenson HB, Sanders WB. Corynebacterium diphtheriae Nelson textbook of Paediatrics. 2000;838-842.
Fletcher B, Dan C. Africa. Sir Banister Fletcher’s History of Architecture. Architectural Press. 1996;1466.
Nakao H, Popovic T. Development of a direct PCR for detection of the diphtheria toxin gene. Journal of Clinical Microbiology. 1997;7:1651–1655.
Nakao H, Popovic T. Development of a direct PCR for detection of the diphtheria toxin gene. Journal of Clinical Microbiology. 1997;35:1651-5.
Youwang Y, Jianming D, Yong X, Pong Z. Epidemiological features of an outbreak of diphtheria and its control with diphtheria toxoid immunization. International journal of Epidemiology. 1992;21(4).
Public Health England. Infection report/ immunization. Diphtheria in England and Wales; Health Protection Weekly Report. 2016;10(13).
Elizabeth A, Mothershed R, Pamela K, Cassiday B, Kevin P, Leonard W, Mayer L, Tanja P. Meningitis and special pathogens branch, division of bacterial and mycotic diseases, national center for infectious diseases, centers for disease control and prevention. Atlanta, Georgia. 2000;30333.
Mikhailovich VM, Melnikov MG, Mazurova IK. Application of PCR for detection of toxigenic Corynebacterium diphtheriae strains isolated during the Russian diphtheria epidemic, 1990 through 1994. Journal of Clinical Microbiology. 1994; 33:3061-3.
Pallen MJ, Hay AJ, Puckey LH, Efstratiou A. Polymerase chain reaction for screening clinicalical isolates of corynebacteria for the production of diphtheria toxin. Journal of Clinical Pathology. 1994;47:353-6.
Efstratiou A, Engler KH, De Zoysa A. Diagnosis and epidemiology of diphtheria. Methods in molecularecular medicine. Molecularecular bacteriologyogy, protocols and clinicalical applications. Humana Press. 1998;15:191-212.
Besa NC, Coldiron ME, Bakri A, Raji A, Nsuami MJ, Rousseau C, Hurtado N, Porten K. Article of Diphtheria outbreak with high mortality in northeastern Nigeria; 2011.
Sadoh AE, Sadoh WE. Diphtheria mortality in Nigeria: The need to stock diphtheria antitoxin. African Journal of Clinical and Experimental Microbiology. 2011;12(2):82-85.
Public Health Laboratory Service. Diphtheria acquired during a cruise in the Baltic Sea. Communicable Disease Representattive CDR Wkly. 1997;7:207.
Snyder JW. Media for detection of Corynebacterium diphtheria. 3.11.7. In Garcia L. S. (ed.), Clinical microbiological procedures handbook, 3rd Ed. ASM Press, Washington DC. 2016;1.
World Health Organization. Diphtheria; 2016.
Engler KH, Glushkevich T, Mazurova IK, George RC, Efstratiou A. A modified Elek test for detection of toxigenic corynebacteria in the diagnostic laboratory. Journal of Clinical Microbiology. 1997;35: 495–498.
Randall KH. Biology and molecular epidemiology of diphtheria toxin and the tox gene. Department of Microbiology, University of Colorado Health Sciences Centre, Denver, Colorado. JID 181: S163; 2000.