Identification of phlebotomine sand fly blood meals by real-time PCR
Background: Phlebotomine sand flies are blood-feeding insects of great medical and veterinary significance acting as vectors of Leishmania parasites. Studying the blood-feeding pattern of these insects may help in the understanding of their interactions with potential reservoir hosts of Leishmania p...
Published in: | Parasites & Vectors |
---|---|
Main Authors: | , , , , , , |
Other Authors: | |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
2015
|
Subjects: | |
Online Access: | http://hdl.handle.net/11586/223441 https://doi.org/10.1186/s13071-015-0840-3 http://www.parasitesandvectors.com/ |
id |
ftunivbari:oai:ricerca.uniba.it:11586/223441 |
---|---|
record_format |
openpolar |
spelling |
ftunivbari:oai:ricerca.uniba.it:11586/223441 2024-02-11T10:08:15+01:00 Identification of phlebotomine sand fly blood meals by real-time PCR Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemos De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemo De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe 2015 http://hdl.handle.net/11586/223441 https://doi.org/10.1186/s13071-015-0840-3 http://www.parasitesandvectors.com/ eng eng info:eu-repo/semantics/altIdentifier/pmid/25889289 info:eu-repo/semantics/altIdentifier/wos/WOS:000353618400001 volume:8 issue:1 Article Number: 230 firstpage:1 lastpage:6 numberofpages:6 journal:PARASITES & VECTORS http://hdl.handle.net/11586/223441 doi:10.1186/s13071-015-0840-3 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84928574239 http://www.parasitesandvectors.com/ info:eu-repo/semantics/openAccess Phlebotomine sand flies Blood meal Brazil Real time PCR info:eu-repo/semantics/article 2015 ftunivbari https://doi.org/10.1186/s13071-015-0840-3 2024-01-17T17:41:05Z Background: Phlebotomine sand flies are blood-feeding insects of great medical and veterinary significance acting as vectors of Leishmania parasites. Studying the blood-feeding pattern of these insects may help in the understanding of their interactions with potential reservoir hosts of Leishmania parasites. In this study, we developed real time PCR assays for the identification of sand fly blood meal. Methods: Six pairs of primers were designed based on cytochrome b gene sequences available in GenBank of the following potential hosts: dog, cat, horse, chicken, black rat, and human.Firstly, SYBR Green-based real time PCR assays were conducted using a standard curve with eight different concentrations (i.e., 10 ng, 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg and 1 fg per 2 μl) of DNA samples extracted from EDTA blood samples from each target animal. Then, DNA samples extracted from field-collected engorged female sand flies belonging to three species (i.e., Lutzomyia longipalpis, L. migonei and L. lenti) were tested by the protocols standardized herein. Additionally, female sand flies were experimentally fed on a black rat (Rattus rattus) and used for evaluating the time course of the detection of the protocol targeting this species. Results: The protocols performed well with detection limits of 10 pg to 100 fg. Field-collected female sand flies were fed on blood from humans (73%), chickens (23%), dogs (22%), horses (15%), black rats (11%) and cats (2%). Interestingly, 76.1% of the L. longipalpis females were positive for human blood. In total, 48% of the tested females were fed on single sources, 31% on two and 12% on three. The analysis of the time course showed that the real time PCR protocol targeting the black rat DNA was able to detect small amounts of the host DNA up to 5 days after the blood meal. Conclusions: The real time PCR assays standardized herein successfully detected small amounts of host DNA in female sand flies fed on different vertebrate species and, specifically for the black rats, up to 5 days ... Article in Journal/Newspaper Rattus rattus Università degli Studi di Bari Aldo Moro: CINECA IRIS Parasites & Vectors 8 1 |
institution |
Open Polar |
collection |
Università degli Studi di Bari Aldo Moro: CINECA IRIS |
op_collection_id |
ftunivbari |
language |
English |
topic |
Phlebotomine sand flies Blood meal Brazil Real time PCR |
spellingShingle |
Phlebotomine sand flies Blood meal Brazil Real time PCR Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemos De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe Identification of phlebotomine sand fly blood meals by real-time PCR |
topic_facet |
Phlebotomine sand flies Blood meal Brazil Real time PCR |
description |
Background: Phlebotomine sand flies are blood-feeding insects of great medical and veterinary significance acting as vectors of Leishmania parasites. Studying the blood-feeding pattern of these insects may help in the understanding of their interactions with potential reservoir hosts of Leishmania parasites. In this study, we developed real time PCR assays for the identification of sand fly blood meal. Methods: Six pairs of primers were designed based on cytochrome b gene sequences available in GenBank of the following potential hosts: dog, cat, horse, chicken, black rat, and human.Firstly, SYBR Green-based real time PCR assays were conducted using a standard curve with eight different concentrations (i.e., 10 ng, 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg and 1 fg per 2 μl) of DNA samples extracted from EDTA blood samples from each target animal. Then, DNA samples extracted from field-collected engorged female sand flies belonging to three species (i.e., Lutzomyia longipalpis, L. migonei and L. lenti) were tested by the protocols standardized herein. Additionally, female sand flies were experimentally fed on a black rat (Rattus rattus) and used for evaluating the time course of the detection of the protocol targeting this species. Results: The protocols performed well with detection limits of 10 pg to 100 fg. Field-collected female sand flies were fed on blood from humans (73%), chickens (23%), dogs (22%), horses (15%), black rats (11%) and cats (2%). Interestingly, 76.1% of the L. longipalpis females were positive for human blood. In total, 48% of the tested females were fed on single sources, 31% on two and 12% on three. The analysis of the time course showed that the real time PCR protocol targeting the black rat DNA was able to detect small amounts of the host DNA up to 5 days after the blood meal. Conclusions: The real time PCR assays standardized herein successfully detected small amounts of host DNA in female sand flies fed on different vertebrate species and, specifically for the black rats, up to 5 days ... |
author2 |
Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemo De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe |
format |
Article in Journal/Newspaper |
author |
Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemos De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe |
author_facet |
Sales, Kamila Gaudêncio Da Silva Costa, Pietra Lemos De Morais, Rayana Carla Silva Otranto, Domenico Brandão-Filho, Sinval Pinto Cavalcanti, Milena De Paiva Dantas-Torres, Filipe |
author_sort |
Sales, Kamila Gaudêncio Da Silva |
title |
Identification of phlebotomine sand fly blood meals by real-time PCR |
title_short |
Identification of phlebotomine sand fly blood meals by real-time PCR |
title_full |
Identification of phlebotomine sand fly blood meals by real-time PCR |
title_fullStr |
Identification of phlebotomine sand fly blood meals by real-time PCR |
title_full_unstemmed |
Identification of phlebotomine sand fly blood meals by real-time PCR |
title_sort |
identification of phlebotomine sand fly blood meals by real-time pcr |
publishDate |
2015 |
url |
http://hdl.handle.net/11586/223441 https://doi.org/10.1186/s13071-015-0840-3 http://www.parasitesandvectors.com/ |
genre |
Rattus rattus |
genre_facet |
Rattus rattus |
op_relation |
info:eu-repo/semantics/altIdentifier/pmid/25889289 info:eu-repo/semantics/altIdentifier/wos/WOS:000353618400001 volume:8 issue:1 Article Number: 230 firstpage:1 lastpage:6 numberofpages:6 journal:PARASITES & VECTORS http://hdl.handle.net/11586/223441 doi:10.1186/s13071-015-0840-3 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84928574239 http://www.parasitesandvectors.com/ |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1186/s13071-015-0840-3 |
container_title |
Parasites & Vectors |
container_volume |
8 |
container_issue |
1 |
_version_ |
1790607278775205888 |