Abstract:
Whipworm infection or trichuriasis caused by Trichuris trichiura is of major public health
concern in sub-Saharan Africa, particularly among pre-school and school-going children.
It is among the neglected tropical diseases targeted for elimination through mass drug
administration (MDA). One of the outcomes of MDA is a rapid decline in levels of infection intensity, making it difficult to monitor effectiveness of control measures using the
conventional Kato–Katz procedure, which relies on the microscopic detection of parasite
ova in faecal samples. In the present study, a loop-mediated isothermal amplification
(LAMP) test was developed for the detection of T. trichiura infection in faecal samples.
LAMP technology offers greater sensitivity and specificity than the microscopy-based
tests. A set of four specific primers targeting the internal transcribed spacer 2 region of
the ribosomal DNA were designed using Primer Explorer software. DNA was extracted
from faecal samples using the alkaline lysis method (HotSHOT) and the LAMP reaction
performed at 63°C for 1 h. The amplicons were visualized by both gel electrophoresis
and with the naked eye following staining with SYBR green dye. Sensitivity and specificity
tests were determined using the standard Kato–Katz diagnostic procedure as a reference
test. The developed LAMP assay reliably detected T. trichiura DNA in faecal samples,
with a specificity and sensitivity of 88% and 77%, respectively. No cross-reactivity was
observed with several common helminth parasites. The developed LAMP assay is an appropriate diagnostic method for the detection of T. trichiura DNA in human faecal samples
due to its simplicity, low cost, high sensitivity and specificity.
