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African swine fever (ASF) is a contagious viral disease, which can cause up to 100% mortality among domestic pigs. Pig production is growing rapidly in Uganda among East African countries and is not only a source of food but also an important income for many people living in the rural areas. Field diagnosis of ASF depends only on clinical signs and has to be confirmed in the laboratory since the clinical signs are not pathognomonic. Diagnostic techniques for ASF are focused on serological tests for detection of antigen and antibody, genomic DNA detection by polymerase chain reaction (PCR), and on virus isolation and localization in clinical samples. There have been many recent reports of ASF outbreaks in Uganda yet laboratory diagnosis is limited due to the high cost and expertise required. This work reports the evaluation and application of a loop-mediated isothermal amplification (LAMP) test for detecting African swine fever virus (ASFV) DNA based on the topoisomerase II gene. Thirty (30) tissue samples obtained from suspected ASF outbreaks were collected from different regions of Uganda. The tissue samples were found to have lesions consistent with ASF. One hundred and eighty eight (188) additional blood samples were obtained from the abattoir and field surveillance. Six primers targeting the topoisomerase II gene were used. The sensitivity and specificity of LAMP and OIE recommended diagnostic PCR were compared. The LAMP assay is rapid with results obtained within 1 h (45-60 min). The sensitivity of LAMP for the detection of ASFV was 100% (95% CI: 91.78-100) while the specificity was 44% (95% CI: 36.52-51.69). The Kappa statistic for level of agreement between PCR and LAMP test in the detection of ASFV was 23.7% (95% CI: 16.42-30.91). This Kappa value indicated a fair agreement between the two assays. No cross reaction was observed with Porcine circovirus type 2 virus and E. coli isolated from pigs in Uganda. This is the first study evaluating and applying the LAMP assay in the detection of ASF in domestic pigs in Uganda. The LAMP assay was found to be more sensitive than PCR. Due to its simplicity, sensitivity and specificity, the LAMP assay has the potential for use in the diagnosis and routine surveillance of ASF in Uganda.       Key words: African swine fever virus, loop-mediated isothermal amplification (LAMP), polymerase chain reaction (PCR), sensitivity, specificity, topoisomerase II gene.

african journal of microbiology research

Comparative detection of African swine fever virus by loop-mediated isothermal amplification assay and polymerase chain reaction in domestic pigs in Uganda