Comparison of the Sensitivities of Different PCR Assays for the Detection of Mycobacterium tuberculosis Complex Isolates from Five Regions of Cameroon

Abstract

The accurate diagnosis of tuberculosis caused by members of the Mycobacterium tuberculosis complex (MTBC) has remained a major challenge in clinical laboratories world-wide. Several studies have evaluated the use of highly specific in-house PCR assays targeting the IS6110, hupB, rpoB, oxyR, and IS1081 genes in the detection of MTBC species with reports on variable sensitivities depending on the geographical sourcing of isolates. In the present investigations, we evaluated the sensitivities of these PCR assays on 125 MTBC cultured isolates from five (West, Centre, Littoral, North West and South West) of the ten regions of Cameroon. Of this number, 124 (99.2%), 117 (93.6%), 123 (99.1%), 119 (95.2%) and 118 (94.4%) were positive by the IS6110, hupB, rpoB, oxyR, and IS1081-based PCR assays respectively. A total of 110 (88%) of the cultured isolates were also identified as MTBC by standard biochemical tests. Of this number, 109 (99.1%), 104 (94.5%), 109 (99.1%), 106 (96.4%) and 104 (94.5%) were positive in the IS6110, hupB, rpoB, oxyR, and IS1081-based PCR assays respectively. Concordant PCR results were obtained for 108 of the 125 samples. The 15 isolates that were negative biochemically scored sensitivities ranging from 100% (for the IS6110 assay) to 86.7% (for the hupB and oxyR assay). The combination of the IS6110 assay, which turned out to be the most sensitive, and each of the other assays gave 100% sensitivity. We conclude that the combined targeting of the IS6110 and rpoB genes is likely to yield the most sensitive PCR procedure for the diagnosis of MTBC infection in the five regions of Cameroon.