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Am. J. Trop. Med. Hyg., 70(6), 2004, pp. 604-606
Copyright © 2004 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT: A NEW POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM METHOD TO IDENTIFY ANOPHELES ARABIENSIS FROM AN. GAMBIAE AND ITS TWO MOLECULAR FORMS FROM DEGRADED DNA TEMPLATES OR MUSEUM SAMPLES

FEDERICA SANTOLAMAZZA, ALESSANDRA DELLA TORRE, AND ADALGISA CACCONE
Department of Ecology and Evolutionary Biology and Yale Institute for Biospheric Studies, Yale University, New Haven, Connecticut; Sezione di Parassitologia, Dipartimento di Scienze di Sanità Pubblica, Università di Roma La Sapienza, Rome, Italy

 

ABSTRACT

We present a polymerase chain reaction-restriction fragment length polymorphism method to simultaneously distinguish the two Anopheles gambiae M and S molecular forms and Anopheles arabiensis. This method uses different diagnostic sites than previously published methods, and it is based on the amplification of a smaller ribosomal DNA fragment. We have tested this protocol in a variety of samples from different geographic regions and various ages of preservation to ascertain the robustness of this protocol over a wide geographic window and on DNA templates of poor quality. This procedure is as efficient as previous ones in discriminating An. arabiensis from the two taxa in An gambiae s.s. However, it performs better than others on poor quality templates such as the ones from museum collections, and poorly stored field collected material. However, it must be noted that it does not allow the simultaneous discrimination of all the species in the An. gambiae complex.

Received November 26, 2003. Accepted for publication January 22, 2004.

Acknowledgments: We thank all the scientists who made this study possible by providing us with field samples, Melissa Garren for laboratory assistance, Mario Coluzzi for stimulating the development of the method, and Jeffrey Powell for financial advice and support. We also thank two anonymous reviewers for their useful comments.

Financial support: Federica Santolamazza and Alessandra Della Torre were supported by the UNDP/World Bank/World Health Organization Special Program for Research and Training in Tropical Diseases (TDR) and by MIUR/COFIN grants. Federica Santolamazza was supported by RO1AI-46018 to Jeffrey Powell while at Yale Univesity.

Authors’ addresses: Federica Santolamazza and Alessandra della Torre, Sezione di Parassitologia, Dipartimento di Scienze di Sanità Pubblica, Università di Roma La Sapienza, Piazzale Aldo Moro, 5 00185 Rome, Italy, Telephone: 39-06-4991-4932, Fax: 39-06-4991-4653, E-mails: Federica.Santolamazza{at}uniroma1.it and ale.dellatorre{at}uniroma1.it. Adalgisa Caccone Yale Institute for Biospheric Studies-Ecosave Molecular Systematics and Conservation Genetics Laboratory, Yale University, 21 Sachem Street, New Haven, CT 06520, Telephone: 203-432-5259 Fax: 203-432-7394, E-mail: adalgisa.caccone{at}yale.edu.




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Copyright © 2004 by the American Society of Tropical Medicine and Hygiene.