A Simple PCR Assay for Discrimination of Dengue Vectors from Nsukka LGA, Nigeria


  • Amos Watentena Department of Zoology and Environmental Biology, University of Nigeria https://orcid.org/0000-0002-9318-9716
  • Ikem Chris Okoye Department of Zoology and Environmental Biology, University of Nigeria
  • Ikechukwu Eugene Onah Department of Zoology and Environmental Biology, University of Nigeria https://orcid.org/0000-0002-2101-7894
  • Onwude Cosmas Ogbonnaya Molecular Unit, National Arbovirus and Vectors Research Centre, Federal Ministry of Health, 33 Park Avenue, GRA, PMB 01573, Enugu, Enugu State, Nigeria
  • Emmanuel Ogudu National Arbovirus and Vectors Research Centre, Federal Ministry of Health, 33 Park Avenue, GRA, PMB 01573, Enugu, Enugu State, Nigeria




Mosquitoes of Aedes species are vectors of several arboviral diseases which continue to be a major public health problem in Nigeria. This study among other things, morphologically identified Aedes mosquitoes collected from Nsukka LGA and used an allele specific PCR amplification for discrimination of dengue vectors. Larval sampling, BG-sentinel traps and modified human landing catches were used for mosquito sampling in two selected autonomous communities of Nsukka LGA (Nsukka and Obimo). A total of 124 Aedes mosquitoes consisting of five (5) different species were collected from April to June, 2019 in a cross-sectional study that covered 126 households, under 76 distinct geographical coordinates. Larvae was mainly collected from plastic containers 73% (n=224), metallic containers 14% (n=43), earthen pots 9% (n=29) and used car tyres 3% (n=9), reared to adult stage 69.35% (n=86), and all mosquitoes were identified using standard morphological keys. Five (5) Aedes mosquito species were captured; Aedes aegypti 83(66.94%), Aedes albopictus 33(26.61%), Aedes simpsoni (4.48%), Aedes luteocephalus (≤1%) and Aedes vittatus (≤1%). Nsukka autonomous community had higher species diversity than Obimo. Allele specific amplification confirmed dengue vectors, Aedes aegypti and Aedes albopictus species on a 2% agarose gel. Since the most recent re-emergence of arboviral diseases is closely associated with Aedes species, findings of this study, therefore, give further evidence about the presence of potential arboviral vectors in Nigeria and describe the role of a simple PCR in discriminating some. Further entomological studies should integrate PCR assays in mosquito vector surveillance.


Aedes aegypti, Aedes albopictus, Dengue


Download data is not yet available.


Malaria Consortium, “Vector control: the untapped potential for neglected tropical diseases,” no. September. Leonard Street, London EC2A4LT, United Kingdom, pp. 1–8, 2017.

J. A. Patz, A. K. Githeko, J. P. McCarty, S. Hussein, U. Confalonieri, and N. de Wet, “Climate change and infectious diseases,” World Health Organization, 2003, pp. 103–132.

A. Watentena, I. Okoye, and I. Onah, “Dengue Reemergence: The Challenges Ahead,” Int. Ann. Sci., vol. 9, no. 1 SE-Review Article, Jun. 2020, doi: 10.21467/ias.9.1.132-140.

B. N. Patrick et al., “Distribution and diversity of mosquitoes and the role of Aedes in the transmission of arboviruses in selected districts of Tanzania,” Int. J. Mosq. Res., vol. 5, no. 1, pp. 53–60, 2018.

L. Braack, A. P. Gouveia De Almeida, A. J. Cornel, R. Swanepoel, and C. De Jager, “Mosquito-borne arboviruses of African origin: Review of key viruses and vectors,” Parasites and Vectors, vol. 11, no. 1, 2018, doi: 10.1186/s13071-017-2559-9.

C. Y. Kow, L. L. Koon, and P. F. Yin, “Detection of dengue viruses in field caught male Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Singapore by type-specific PCR,” J. Med. Entomol., vol. 38, no. 4, pp. 475–479, 2001, doi: 10.1603/0022-2585-38.4.475.

J. A. Jackman and J. K. A. J. and J. K. Olson, “Mosquitoes and the diseases they transmit,” Agric. Commun. Texas A&M Univ. Syst. Ext. Publ., vol. B-6119, pp. 1–8, 2011, doi: 10.1007/s10531-005-2089-1.

WHO/WMO, Health and climate, WMO-No. 10. 20 Avenue Appia, 1211 Geneva 27, Switzerland: WHO Press, 2012.

M. Kashif Zahoor et al., Dengue fever: a general perspective. IntechOpen, 2019.

M. L. Phillips, “Dengue reborn,” Environ. Health Perspect., vol. 116, no. 9, pp. 382–388, 2008, doi: 10.1289/ehp.116-a382.

World Health Organization, “Global strategy for dengue prevention and control 2012-2020,” Geneva, 2012.

M. S. A. Alshehri, “Dengue fever outburst and its relationship with climatic factors,” World Appl. Sci. J., vol. 22, no. 4, pp. 506–515, 2013, doi: 10.5829/idosi.wasj.2013.22.04.443.

Global Malaria Programme, “Tailoring malaria interventions in the COVID-19 response,” Geneva, 2020.

G. Benelli, “Managing mosquitoes and ticks in a rapidly changing world – Facts and trends,” Saudi J. Biol. Sci., vol. 26, no. 5, pp. 921–929, 2019, doi: 10.1016/j.sjbs.2018.06.007.

A. Amarasinghe, J. N. Kuritsky, G. William Letson, and H. S. Margolis, “Dengue virus infection in Africa,” Emerg. Infect. Dis., vol. 17, no. 8, pp. 1349–1354, 2011, doi: 10.3201/eid1708.101515.

F. Were, “The dengue situation in Africa,” Paediatr. Int. Child Health, vol. 32, no. Sup1, pp. 18–21, 2012, doi: 10.1179/2046904712Z.00000000048.

J. Soghigian, T. G. Andreadis, and T. P. Livdahl, “From ground pools to treeholes: convergent evolution of habitat and phenotype in Aedes mosquitoes,” BMC Evol. Biol., vol. 17, no. 1, pp. 1–13, 2017, doi: 10.1186/s12862-017-1092-y.

D. Guha-Sapir and B. Schimmer, “Dengue fever: New paradigms for a changing epidemiology,” Emerg. Themes Epidemiol., vol. 2, no. 1, pp. 1–10, 2005, doi: 10.1186/1742-7622-2-1.

M. Q. Benedict, R. S. Levine, W. A. Hawley, and L. P. Lounibos, “Spread of the Tiger: global risk invasion by the mosquito Aedes albopictus,” Vector Borne Zoonotic Dis., vol. 7, no. 1, pp. 76–85, 2007, doi: 10.1055/s-0029-1237430.Imprinting.

G. Rezza, “Aedes albopictus and the reemergence of dengue,” BMC Public Health, vol. 12, no. 1, pp. 2–4, 2012, doi: 10.1186/1471-2458-12-72.

WHO, “Dengue,” 20 Avenue Appia, 1211 Geneva 27, Switzerland, 2014.

K. J. Linthicum, P. G. Da Vies, and A. Kairo, “Rift Valley fever virus (family Bunyaviridae, genus Phlebovirus). Isolations from Diptera collected during an inter-epizootic period in Kenya,” J. Hyg. (Lond)., vol. 95, no. 1, pp. 197–209, 1985, doi: 10.1017/S0022172400062434.

B. J. Trewin et al., “The elimination of the dengue vector, Aedes aegypti, from Brisbane, Australia: The role of surveillance, larval habitat removal and policy,” PLoS Negl. Trop. Dis., vol. 11, no. 8, pp. 1–23, 2017, doi: 10.1371/journal.pntd.0005848.

G. E. Odo, E. J. Agwu, and A. S. Haruna, “Culicid forms distribution and breeding sites in Nsukka ecological zone of South Eastern Nigeria,” J. Parasitol. Vector Biol., vol. 7, no. 5, pp. 94–100, 2015, doi: 10.5897/JPVB2015.0193.

M. N. Ezike et al., “A preliminary study of mosquito vectors of human diseases and infection rates in Oru West, Imo State. Nigeria,” Glob. Res. J. Sci., vol. 5, no. 1, pp. 1–10, 2017.

C. O. Chimaeze et al., “Diversity and distribution of Aedes mosquitoes in Nigeria,” New York Sci. J., vol. 11, no. 2, 2018.

S. B. Kumar, M. R. Prusty, A. Rath, S. K. Kar, and R. K. Hazra, “A simple, rapid and very efficient protocol for DNA isolation from mosquito species Sushanta.” Regional Medical Research Centre, Nalco Square, Chandrasekharpur, Bhubaneswar 751023, India, pp. 3–8, 2010.

K. S. Walter, J. E. Brown, and J. R. Powell, “Microhabitat partitioning of Aedes simpsoni (Diptera: Culicidae),” J. Med. Entomol., vol. 51, no. 3, pp. 596–604, 2014, doi: 10.1603/me13097.

N. Becker et al., “Medical importance of mosquitoes,” in Mosquitoes and their control, Springer Berlin Heidelberg, 2010, pp. 25–42.

FRNOG, “Legal notice on publication of the details of the breakdown of the national and state provisional totals 2006 census,” 2007. .

Climate Data, “Climate of Nsukka,” 2020. [Online]. Available: https://en.climate-data.org/location/404904/. [Accessed: 10-Jul-2020].

J. D. Gillett, Common African mosquitos and their medical importance. William Heinemann, London, UK, 1972.

E. H. Simpson, “Measurement of diversity,” Nature, vol. 163, no. 4148, p. 688, 1949, doi: 10.1038/163688a0.

S. C. Weaver and W. K. Reisen, “Present and future arboviral threats,” Antiviral Res., vol. 85, no. 2, pp. 328–345, 2010, doi: 10.1016/j.antiviral.2009.10.008.

NCDC, “Response to yellow fever cases in Edo State,” Press Release, 24-Nov-2018. [Online]. Available: https://ncdc.gov.ng/news/157/response-to-yellow-fever-cases-in-edo-state.

A. B. Onoja, J. A. Adeniji, and O. D. Olaleye, “High rate of unrecognized dengue virus infection in parts of the rainforest region of Nigeria,” Acta Trop., vol. 160, pp. 39–43, 2016, doi: 10.1016/j.actatropica.2016.04.007.

C. K. Ezihe et al., “Mosquito species associated with refuse dumps within Enugu Municipal, Enugu State, Nigeria,” Int. J. Mosq. Res., no. February, 2017, doi: 10.5376/jmr.2017.07.0006.

D. N. Aribodor, O. B. Aribodor, E. C. I., and A. E. Onyido, “Survey of open larval habitats of mosquitoes in four communities in Awka South Local Government Area, Anambra State, Nigeria,” Biosci., vol. 1, no. 2, pp. 132–139, 2013.

O. A. E. et al., “Anthropophilic mosquito species prevalence in Nibo Community, Awka South Local Government Area, Anambra State, Southeastern Nigeria,” Ewemen J. Epidemiol. Clin. Med., vol. 2, no. 1, pp. 14–20, 2016.

G. R. A. Okogun, J. C. Anosike, A. N. Okere, and B. E. B. Nwoke, “Ecology of mosquitoes of Midwestern Nigeria,” J. Vector Borne Dis., vol. 42, no. 1, pp. 1–8, 2005.

U. C. Chukwuekezie, O. C. Nwangwu et al., “A Cross sectional survey of yellow fever and dengue virus vectors in four communities of Ayamelum Local Government Area (LGA), Anambra State, Southeast Nigeria,” New York Sci. J., vol. 9, no. 3, 2018, doi: 10.7537/marsnys09031616.

K. L. Bennett et al., “Molecular differentiation of the African yellow fever vector Aedes bromeliae (Diptera: Culicidae) from its sympatric non-vector sister species, Aedes lilii,” PLoS Negl. Trop. Dis., vol. 9, no. 12, pp. 1–19, 2015, doi: 10.1371/journal.pntd.0004250.

Y. Higa, T. Toma, Y. Tsuda, and I. Miyagi, “A multiplex PCR-based molecular identification of five morphologically related, medically important subgenus stegomyia mosquitoes from the genus Aedes (Diptera: Culicidae) found in the Ryukyu Archipelago, Japan,” Jpn. J. Infect. Dis., vol. 63, no. 5, pp. 312–316, 2010.

H. Iwashita et al., “Mosquito arbovirus survey in selected areas of Kenya: detection of insect-specific virus,” Trop. Med. Health, vol. 46, no. 1, pp. 1–15, 2018, doi: 10.1186/s41182-018-0095-8.






Research Article

How to Cite

A. Watentena, I. C. Okoye, I. E. Onah, O. C. Ogbonnaya, and E. Ogudu, “A Simple PCR Assay for Discrimination of Dengue Vectors from Nsukka LGA, Nigeria”, Int. Ann. Sci., vol. 10, no. 1, pp. 67-77, Oct. 2020.