Inter-Simple Sequence Repeat (ISSR) Polymorphism of Tuberculosis Patients from Mile Four Hospital, Abakaliki, Nigeria

Abstract

Tuberculosis (TB) is a serious health issue in Ebonyi State and Nigeria at large. The knowledge of genetic diversity and population structure of TB patients in Ebonyi State is still inadequate. Therefore, this work tends to assess the genetic diversity and population structure in association with tuberculosis population using Inter-simple sequence repeat (ISSR) markers. A total of 25 samples were collected, in which 21 were from TB patients and 4 from NTB patients as controls. DNA extraction was done using Cetyltrimethylammonium bromide (CTAB) method followed by Polymerase chain reaction (PCR) with ISSR primers. Analyses of dendrogram, principal component analysis, genetic diversity and population structure were computed with the scored matrix data. Six ISSR markers were polymorphic. The polymorphic information content (PIC) values ranged from 0.2635 to 0.8158 with a mean of 0.6156. The genetic diversity ranged from 0.1504 to 0.8352 with a mean of 0.6156, while the major allele frequency ranged from 0.2800 to 0.9200 with a mean of 0.5018. The allele count ranged from 1 to 23, while the frequency ranged from 0.0400 to 0.9200. The values of number of alleles (Na), effective number of alleles (Ne), Nei's gene diversity (H) and Shannon's information index (I) from the entire population were 2.000±0.000, 1.8562±0.1011, 0.4595±0.0330 and 0.6517±0.0350, respectively while that of the six ISSR markers ranges from 1.3200-2.0000, 1.1184-1.9091, 0.0456-0.4571 and 0.1388-0.6611, respectively. From the six ISSR marker data, the number of polymorphic loci (NPL) and percentage polymorphic loci (PPL) ranged from 2 to 25 and 32% to 100% respectively. A dendrogram of the 25 TB and non- NTB clustered them into nine major groups. This study revealed high diversity among TB patientsatgene loci also probed, but did not show any clear association between TB susceptibility and the gene loci.