TY - JOUR
T1 - PCR-RFLP method for the identification of four members of the Anopheles annularis group of mosquitoes (Diptera: Culicidae)
AU - Alam, Mohammad Tauqeer
AU - Das, Manoj K.
AU - Dev, Vas
AU - Ansari, Musharraf A.
AU - Sharma, Yagya D.
N1 - Funding Information:
We are grateful to Prof. A.P. Dash (Director, National Institute of Malaria Research, Delhi) for his kind help and support. We also thank Ms Sumiti Vinayak for discussion and critical reading of the manuscript. M.T.A. acknowledges the Council of Scientific and Industrial Research for a Senior Research Fellowship. Financial support for the work granted by the Indian Council of Medical Research, and Department of Biotechnology, Government of India, is acknowledged. We are also grateful to the Bioinformatics facilities of BTIS.
Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 2007/3
Y1 - 2007/3
N2 - The Anopheles annularis group of mosquitoes is widely distributed in Southeast Asia and may be locally important as malaria vectors. Members of this group are morphologically very similar and often difficult to distinguish, particularly A. nivipes and A. philippinensis. We report the sequence analysis of the rDNA internal transcribed spacer 2 (ITS2) and Domain-3 (D3) regions of the four members of the A. annularis group - A. nivipes, A. philippinensis, A. annularis and A. pallidus - and a method for their molecular identification. No intraspecies sequence variation was detected among the specimens, while interspecific sequence differences were greater for ITS2 than the D3 regions. Comparison of the D3 sequences of the four species revealed two SmaI restriction sites in A. nivipes, but only one site in A. philippinensis, A. annularis and A. pallidus. The ApaI site was present in both A. philippinensis and A. pallidus, while an NcoI site was present in A. pallidus only. Restriction digestion of the PCR products of D3 fragment individually with SmaI, ApaI and NcoI produced a distinctive pattern for all the four species. We present, for the first time, a PCR-RFLP method to distinguish the four members of the A. annularis group of mosquitoes.
AB - The Anopheles annularis group of mosquitoes is widely distributed in Southeast Asia and may be locally important as malaria vectors. Members of this group are morphologically very similar and often difficult to distinguish, particularly A. nivipes and A. philippinensis. We report the sequence analysis of the rDNA internal transcribed spacer 2 (ITS2) and Domain-3 (D3) regions of the four members of the A. annularis group - A. nivipes, A. philippinensis, A. annularis and A. pallidus - and a method for their molecular identification. No intraspecies sequence variation was detected among the specimens, while interspecific sequence differences were greater for ITS2 than the D3 regions. Comparison of the D3 sequences of the four species revealed two SmaI restriction sites in A. nivipes, but only one site in A. philippinensis, A. annularis and A. pallidus. The ApaI site was present in both A. philippinensis and A. pallidus, while an NcoI site was present in A. pallidus only. Restriction digestion of the PCR products of D3 fragment individually with SmaI, ApaI and NcoI produced a distinctive pattern for all the four species. We present, for the first time, a PCR-RFLP method to distinguish the four members of the A. annularis group of mosquitoes.
KW - Anopheles annularis
KW - Anopheles nivipes
KW - Anopheles pallidus
KW - Anopheles philippinensis
KW - D3 domain
KW - PCR-RFLP
KW - rDNA ITS2
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U2 - 10.1016/j.trstmh.2006.03.007
DO - 10.1016/j.trstmh.2006.03.007
M3 - Article
C2 - 16806334
AN - SCOPUS:33846012549
SN - 0035-9203
VL - 101
SP - 239
EP - 244
JO - Transactions of the Royal Society of Tropical Medicine and Hygiene
JF - Transactions of the Royal Society of Tropical Medicine and Hygiene
IS - 3
ER -