Skip to main content

Advertisement

Log in

Detection of Onchocerca volvulus in Latin American black flies for pool screening PCR using high-throughput automated DNA isolation for transmission surveillance

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Basáñez MG, Boussinesq M, Prod’hon J, Frontado H, Villamizar NJ, Medley GF, Anderson RM (1994) Density-dependent processes in the transmission of human onchocerciasis: intensity of microfilariae in the skin and their uptake by the simuliid host. Parasitology 108:115–127

    Article  PubMed  Google Scholar 

  • Basáñez MG, Razalik, Renz A, Kelly D (2007) Density-dependent host choice by disease vectors epidemiological implications of the ideal free distribution. Trans R Soc Trop Med Hyg 101:256–269

    Article  PubMed  Google Scholar 

  • Bextine B, Tuan SJ, Sheikh H, Blua M, Miller TA (2004) Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy winged sharpshooter. J Econ Entomol 97:757–763

    Article  PubMed  CAS  Google Scholar 

  • Chan PKS, Chan DPC, To KF, Yu MY, Cheung JLK, Cheng AF (2001) Evaluation of extraction methods from paraffin wax embedded tissues for PCR amplification of human and viral DNA. J Clin Pathology 54:401–403

    Article  CAS  Google Scholar 

  • Chen H, Rangasamy M, Tan SY, Wang H, Siegfried BD (2010) Evaluation of five methods for total DNA extraction from Western corn rootworm beetles. PLoS ONE 5(8): e11963

    Google Scholar 

  • Davies JB, Oskam L, Lujan R, Schoone GH, Kroon CCM, Lopez-Martinez LA, Paniagua-Alvarez AJ (1998) Detection of Onchocerca volvulus DNA in pools of wild caught Simulium ochraceum by use of the polymerase chain reaction. Ann Trop Med Parasitol 92:295–304

    Article  PubMed  CAS  Google Scholar 

  • Figueroa MH, Collins C, Kozek WJ (1977) Post-prandial transportation and maintenance of Simulium ochraceum infected with Onchocerca volvulus. Am J Trop Med Hyg 26:75–79

    Google Scholar 

  • Gonzalez RJ, Ortiz NC, Rizzo N, Richards J, Flores GZ, Domínguez A, Sauerbrey M, Catú E, Oliva O, Richards FOJ, Lindblade KA (2009) Successful interruption of transmission of Onchocerca volvulus in the Escuintla-Guatemala focus, Guatemala. PLoS Negl Trop Dis 3(3):e404

    Google Scholar 

  • Gopal H, Hassan KH, Rodríguez-Pérez MA, Toé DL, Lustigman S, Unnasch TR (2012) Oligonucleotide-based magnetic bead capture of Onchocerca volvulus DNA for pool screening of vector black flies. PLoS Negl Trop Dis 6(6):1712

    Article  Google Scholar 

  • Katholi KR, Toé L, Merriweather A, Unnasch TR (1995) Determining the prevalence of Onchocerca volvulus infection in vector populations by screening pools of blackflies. J Infect Dis 172:1414–1417

    Article  PubMed  CAS  Google Scholar 

  • Korin BP (1977) Introduction to Statistical Methods. Winthrop Publishers, Inc., Cambridge, United Kingdom

    Google Scholar 

  • Meredith SEO, Lando G, Gbakima AA, Zimmerman PA, Unnasch TR (1991) Onchocerca volvulus: application of the polymerase chain reaction to identification and strain differentiation of the parasite. Exp Parasitol 73:335–344

    Article  PubMed  CAS  Google Scholar 

  • Merriweather A, Unnasch TR (1996) Onchocerca volvulus: development of species-specific polymerase chain reaction-based assay. Exp Parasitol 83:164–166

    Article  PubMed  CAS  Google Scholar 

  • Rådström P, Knutsson R, Wolffs J, Lövenklev M, Löfström C (2004) Pre-PCR processing: strategies to generate PCR-compatible samples. Mol Biotechnol 26:133–146

    Article  PubMed  Google Scholar 

  • Reineke A, Karlovsky P, Zebitz CPW (1998) Preparation and purification of DNA from insects for AFLP analysis. Insect Mol Biol 7:95–99

    Article  PubMed  CAS  Google Scholar 

  • Rodríguez-Pérez MA, Reyes-Villanueva F (1994) The effect of ivermectin on the transmission of Onchocerca volvulus in southern Mexico. Salud Publica Mex 36:281–290

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Rodríguez MH, Margeli-Pérez HM, Rivas-Alcalá AR (1995) Effect of semiannual treatments of ivermectin on the prevalence and intensity of Onchocerca volvulus skin infection, ocular lesions, and on the infectivity of Simulium ochraceum populations in Southern Mexico. Am J Trop Med Hyg 52:429–434

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Danis-Lozano R, Rodríguez MH, Bradley JE (1999) Comparison of serological and parasitological assessments of Onchocerca volvulus transmission after 7 years of mass ivermectin treatment in Mexico. Trop Med Intl Health 4:98–104

    Article  Google Scholar 

  • Rodríguez-Pérez MA, Lilley BG, Domínguez-Vázquez A, Segura-Arenas R, Lizarazo-Ortega C, Mendoza-Herrera A, Reyes-Villanueva F, Unnasch TR (2004) Polymerase chain reaction monitoring of transmission of Onchocerca volvulus in two endemic states in Mexico. Am J Trop Med Hyg 70:38–45

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Katholi CR, Hassan HK, Unnasch TR (2006) Large-scale entomologic assessment of Onchocerca volvulus transmission by poolscreen PCR in Mexico. Am J Trop Med Hyg 74:1026–1033

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Segura-Cabrera A, Lizarazo-Ortega C, Maria-Gloria B, Davis JB (2007) Contribution of migrant coffee laborers infected with Onchocerca volvulus to the maintenance of the microfilarial reservoir in an ivermectin-treated area of Mexico. Filarial Journal 6:16–26

    Article  Google Scholar 

  • Rodríguez-Pérez MA, Lutzow-Steiner MA, Segura-Cabrera A, Lizarazo-Ortega A, Domínguez-Vázquez A, Sauerbrey M, Richards F Jr, Unnasch TR, Hassan HK, Hernández-Hernández R (2008a) Rapid suppression of Onchocerca volvulus transmission in two communities of the Southern Chiapas Focus, Mexico, achieved by quarterly treatments with Mectizan. Am J Trop Med Hyg 79:239–244

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Ortega CL, Hassan HK, Vásquez AD, Galván JM, Moreno PL, Sauerbrey M, Richards F Jr, Unnasch TR (2008b) Evidence for suppression of Onchocerca volvulus transmission in the Oaxaca Focus in Mexico. Am J Trop Med Hyg 78:147–152

    PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Unnasch TR, Domínguez-Vázquez A, Morales-Casta AL, Peña-Flores GP, Algarra-Orozco ME, Arredondo-Jiménez JI, Richards F Jr, Rodríguez-V MA, Rendón VG (2010a) Interruption of transmission of Onchocerca volvulus in the Oaxaca focus, Mexico. Am J Trop Med Hyg 83:21–27

    Article  PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Unnasch TR, Domínguez-Vázquez A, Morales-Casta AL, Richards F Jr, Miguel A, Rodríguez VP, Peña-Flores, Prado-Velasco G (2010b) Lack of active Onchocerca volvulus transmission in the Northern Chiapas focus in Mexico. Am J Trop Med Hyg 83:15–20

    Article  PubMed  Google Scholar 

  • Rodríguez-Pérez MA, Unnasch TR, Real-Najarro O (2011) Assessment and monitoring of onchocerciasis in Latin America. Adv Parasitol 77:175–226

    Article  PubMed  Google Scholar 

  • Vieira JC, Cooper PJ, Lovato R, Mancero T, Rivera J, Proaño R, López AA, Guderian RH, Guzmán JR (2007) Impact of long-term treatment of onchocerciasis with ivermectin in Ecuador: potential for elimination of infection. BMC Medicine 5:1–10

    Article  Google Scholar 

  • WER (2012) Progress towards eliminating onchocerciasis in the WHO Region of the Americas in 2011: interruption of transmission in Guatemala and Mexico. Wkly Epidemiol Rec 87(33):305–316

    Google Scholar 

  • Wiedbrauk DL, Werner JC, Drevon AM (1995) Inhibition of PCR by aqueous and vitreous fluids. J Clinical Microbiol 33:2643–2646

    CAS  Google Scholar 

  • World Health Organization (2001) Certification of elimination of human onchocerciasis: criteria and procedures. Geneva, Switzerland, World Health Organization. Document WHO/CDS/CPE/CEE’2001.

Download references

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).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mario A. Rodríguez-Pérez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rodríguez-Pérez, M.A., Gopal, H., Adeleke, M.A. et al. Detection of Onchocerca volvulus in Latin American black flies for pool screening PCR using high-throughput automated DNA isolation for transmission surveillance. Parasitol Res 112, 3925–3931 (2013). https://doi.org/10.1007/s00436-013-3583-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-013-3583-0

Keywords

Navigation