TY - JOUR
T1 - Detection of Onchocerca volvulus in Latin American black flies for pool screening PCR using high-throughput automated DNA isolation for transmission surveillance
AU - Rodríguez-Pérez, Mario A.
AU - Gopal, Hemavathi
AU - Adeleke, Monsuru Adebayo
AU - De Luna-Santillana, Erick Jesús
AU - Gurrola-Reyes, J. Natividad
AU - Guo, Xianwu
N1 - Funding Information:
Acknowledgments We thank Dr. Thomas R. Unnasch (University of South Florida, Tampa Florida, USA) who critically reviewed an earlier version of the manuscript. We would like to thank Promega Corporation for providing us the Maxwell 16 Instrument. We are very grateful to the people from Las Golondrinas in the Southern Chiapas focus who enthusiastically participated throughout this study. We thank MSc Cristian Lizarazo-Ortega, from our Graduate Department, for assisting and helping during the initial phase of the laboratory work. Mario A. Rodríguez-Pérez and Xianwu Guo hold a scholarship from Comisión de Operación y Fomento de Actividades Académicas (COFAA)/Instituto Politécnico Nacional (IPN). Hemavathi Gopal thanks the Consejo Nacional de Ciencia y Tecnología-México (CONACYT) for providing a doctoral scholarship (reference no. 7063). This study was financially supported by CONACYT–México (grant no. 874080).
PY - 2013/11
Y1 - 2013/11
N2 - The posttreatment entomological surveillance (ES) of onchocerciasis in Latin America requires quite large numbers of flies to be examined for parasite infection to prove that the control strategies have worked and that the infection is on the path of elimination. Here, we report a high-throughput automated DNA isolation of Onchocerca volvulus for PCR using a major Latin American black fly vector of onchocerciasis. The sensitivity and relative effectiveness of silica-coated paramagnetic beads was evaluated in comparison with phenol chloroform (PC) method which is known as the gold standard of DNA extraction for ES in Latin America. The automated method was optimized in the laboratory and validated in the field to detect parasite DNA in Simulium ochraceum sensu lato flies in comparison with PC. The optimization of the automated method showed that it is sensitive to detect O. volvulus with a pool size of 100 flies as compared with PC which utilizes 50 flies pool size. The validation of the automated method in comparison with PC in an endemic community showed that 5/67 and 3/134 heads pools were positive for the two methods, respectively. There was no statistical variation (P < 0.05) in the estimation of transmission indices generated by automated method when compared with PC method. The fact that the automated method is sensitive to pool size up to 100 confers advantage over PC method and can, therefore, be employed in large-scale ES of onchocerciasis transmission in endemic areas of Latin America.
AB - The posttreatment entomological surveillance (ES) of onchocerciasis in Latin America requires quite large numbers of flies to be examined for parasite infection to prove that the control strategies have worked and that the infection is on the path of elimination. Here, we report a high-throughput automated DNA isolation of Onchocerca volvulus for PCR using a major Latin American black fly vector of onchocerciasis. The sensitivity and relative effectiveness of silica-coated paramagnetic beads was evaluated in comparison with phenol chloroform (PC) method which is known as the gold standard of DNA extraction for ES in Latin America. The automated method was optimized in the laboratory and validated in the field to detect parasite DNA in Simulium ochraceum sensu lato flies in comparison with PC. The optimization of the automated method showed that it is sensitive to detect O. volvulus with a pool size of 100 flies as compared with PC which utilizes 50 flies pool size. The validation of the automated method in comparison with PC in an endemic community showed that 5/67 and 3/134 heads pools were positive for the two methods, respectively. There was no statistical variation (P < 0.05) in the estimation of transmission indices generated by automated method when compared with PC method. The fact that the automated method is sensitive to pool size up to 100 confers advantage over PC method and can, therefore, be employed in large-scale ES of onchocerciasis transmission in endemic areas of Latin America.
UR - http://www.scopus.com/inward/record.url?scp=84887239474&partnerID=8YFLogxK
U2 - 10.1007/s00436-013-3583-0
DO - 10.1007/s00436-013-3583-0
M3 - Artículo
SN - 0932-0113
VL - 112
SP - 3925
EP - 3931
JO - Parasitology Research
JF - Parasitology Research
IS - 11
ER -