Skip to main content

Advertisement

SpringerLink
Log in

Use of a simple DNA extraction method for high-throughput detection of filarial parasite Wuchereria bancrofti in the vector mosquitoes

  • Short Communication
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Molecular xenomonitoring of filariasis is the detection of filarial DNA in mosquitoes by PCR and a useful tool for monitoring transmission. DNA extraction coupled with PCR allows rapid detection of the presence or absence of the filarial parasite in vector mosquitoes compared to traditional method of manual dissection of the mosquito and observation for parasite under a microscope. A Tris–EDTA (TE) buffer-based boiling method of DNA extraction developed earlier by us was employed and explored for its suitability in the detection of Wuchereria bancrofti DNA in pools of Culex quinquefasciatus mosquitoes in real-time PCR assay. In this preliminary study, 1,000 laboratory-reared C. quinquefasciatus were made into 40 pools, each containing 25 mosquitoes spiked with 2mf. DNA from the first 20 pools was extracted using Qiagen DNeasy blood and tissue kit as standard, and the other 20 pools were subjected to TE buffer-based boiling method of DNA extraction. When the results (Ct values) obtained for DNA samples extracted by TE buffer-based boiling method were compared with that of the DNA samples extracted by the standard Qiagen method, they were found to be highly concordant without any significant difference (P = 0.9). Besides being cost- and time-effective, this protocol was found useful in extracting filarial DNA from two other mosquito genus Aedes and Anopheles, species of which have been reported as important vectors of W. bancrofti in other endemic regions of the world. Thus, TE buffer-based boiling method of DNA extraction is useful for the high-throughput detection of W. bancrofti in vector mosquitoes.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

References

  • Chandrashekar R, Rao UR, Rajasekariah GR, Subrahmanyam D (1984) Separation of viable microfilariae free of blood cells on Percoll gradients. J Helminthol 58:69–70

    Article  PubMed  CAS  Google Scholar 

  • Chanteau S, Luquiaud P, Failloux AB, Williams SA (1994) Detection of Wuchereria bancrofti larvae in pools of mosquitoes by the polymerase chain reaction. Trans R Soc Trop Med Hyg 88:665–666

    Article  PubMed  CAS  Google Scholar 

  • Farid HA, Zakariya SM, Helmy H, Ramzy RMR, El Setouby M, Weil GJ (2007) A critical appraisal of molecular xenomonitoring as a tool for assessing progress toward elimination of lymphatic filariasis. Am J Trop Med Hyg 77:599–600

    Google Scholar 

  • Ottesen EA (2000) The global programme to eliminate lymphatic filariasis. Trop Med Int Health 5:591–594

    Article  PubMed  CAS  Google Scholar 

  • Ramzy RMR, Farid HA, Kamal IH, Ibrahim GH, Morsey ZS, Faris R, Weil GJ, Williams SA (1997) A polymerase chain reaction-based assay for detection of Wuchereria bancrofti in human blood and Culex pipiens. Trans R Soc Trop Med Hyg 9:156–160

    Article  Google Scholar 

  • Rao RU, Atkinson LJ, Ramzy RMR, Helmy H, Farid HA, Bockarie MJ, Susapu M, Laney SJ, Williams SA, Weil GJ (2006) A real-time PCR based assay for detection of Wuchereria bancrofti DNA in blood and mosquitoes. Am J Trop Med Hyg 74:826–832

    PubMed  CAS  Google Scholar 

  • Rocha E, Fontes G, Almeida WAP (2005) Validation of tools for verification of elimination of transmission. 2004–2005; PCR and Xenomonitoring Results from Belém and Maceió, Brazil. 5th RPRG. Meeting Report. www.paho.org/English/AD/DPC/CD/PMM3d-Cntry-Pres4of5.pdf. Accessed 28 Dec 2007

  • Vasuki V, Patra KP, Hoti SL (2001) A rapid and simplified method of DNA extraction for the detection of Brugia malayi infection in mosquitoes by PCR assay. Acta Trop 79:245–248

    Article  PubMed  CAS  Google Scholar 

  • Vasuki V, Hoti SL, Sadanandane C, Jambulingam P (2003) A simple and rapid extraction method for the detection of Wuchereria bancrofti infection in vector mosquito, Culex quinquefasciatus by Ssp 1PCR assay. Acta Trop 86:109–114

    Article  PubMed  CAS  Google Scholar 

  • WHO (2006) Global programme to eliminate lymphatic filariasis. WHO Wkly Epidemiol Rec 81(22):221–232

    Google Scholar 

  • Zhong M, McCarthy J, Bierwert L, Lizotte-Waniewski M, Chanteau S, Nutman TB, Ottesan E, Williams SA (1996) A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples. Am J Trop Med Hyg 54:357–363

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors are thankful to R. Balasubramanian and K. Nagalakshmi for their technical assistance.

Author information

Authors and Affiliations

  1. Vector Control Research Centre (ICMR), Indra Nagar, Medical Complex, Puducherry, 605 006, India

    V. Vasuki, S. Subramanian, S. L. Hoti & P. Jambulingam

Authors
  1. V. Vasuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Subramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. L. Hoti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. Jambulingam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. Vasuki.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vasuki, V., Subramanian, S., Hoti, S.L. et al. Use of a simple DNA extraction method for high-throughput detection of filarial parasite Wuchereria bancrofti in the vector mosquitoes. Parasitol Res 111, 2479–2481 (2012). https://doi.org/10.1007/s00436-012-3026-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-012-3026-3

Keywords

Search

Navigation