Suitability of a rapid DNA isolation and amplification for detection ofTrypanosoma cruziinTriatoma infestansdry fecal spots collected on filter paper

Abstract

Summary :A rapid DNA extraction was used for T. cruzi detection in triatomines dry fecal spots collected on filter paper and analyzed by PCR. Fifty T. infestans were fed on experimentally infected Balb/C mice with high T. cruzi parasitemia and divided into five groups of ten triatomines, and 100 triatomines were infected with lower parasitemia and divided into five groups of 20 triatomines. One dry fecal spot was analyzed per group on days 1, 2, 3, 4 and 5 post feeding. Amplification targeted T. cruzi TCZ sequence and resulted positive from day 4 after bugs feeding in the two models (high and lower parasitemia). The rapid DNA isolation and PCR proposed are suitable for detection of T. cruzi DNA in filter paper and should be considered in field research.

Target Product Profile (TPP) for Chagas Disease Point-of-Care Diagnosis and Assessment of Response to Treatment

“…For other settings, more complex technologies such as enzyme-linked immunosorbent assay (ELISA) and PCR might be acceptable. Molecular methods applied to dried blood on filter paper, which has been carried out for triatomine samples [ 28 , 29 ], could enhance detection accuracy in acute and congenital Chagas disease cases. Moreover, serological methods based on low-volume samples are currently under standardization and validation [ 30 ].…”

Bias due to methods of parasite detection when estimating prevalence of infection ofTriatoma infestansbyTrypanosoma cruzi

“…PCR has already been found to be more sensitive than microscopy for the detection of the parasite, both in blood samples of patients and in triatomines (Botto‐Mahan et al , Braz et al , Dorn et al , Dorn et al , Kirchhoff et al , Pizarro et al ). Dead insect or dry material can be PCR processed, facilitating field handling of samples (Braz et al , Russomando et al ). The DNA extraction protocol used in the present study was carried out on rectal gland tissues from insect dissections, avoiding taking the stomach, a source of PCR inhibitors (Dorn et al ).…”

Dried Blood Spots technology for veterinary applications and biological investigations: technical aspects, retrospective analysis, ongoing status and future perspectives

“…Specific tasks in veterinary and biological research also employed a wide range of dried biosamples such as milk (Wu et al 2008 ; Samsonova et al 2014 , 2017 ; Durel et al 2015 ; Venkatesh and Gopal 2018 ), serosanguineous fluid from the throracic cavity (Elmore et al 2014 ), spleen and lymph node aspirates (Strauss-Ayali et al 2004 ), tongue epithelial (Muthukrishnan et al 2008 ; Madhanmohan et al 2013 , 2016 ; Biswal et al 2016 ) and foot epithelium samples (Madhanmohan et al 2016 ), brain tissue spots (Wacharapluesadee et al 2003 ; Jefferies et al 2007 ; Sakai et al 2015 ; Léchenne et al 2016 ; Rasolonjatovo et al 2020 ), bone marrow aspirates (Cortes et al 2004 ), bovine semen (Sarangi et al 2018 ), haemolymph (Machado et al 2000 ; Kiatpathomchai et al 2004 ; Sudhakaran et al 2009 ), fish body mucus and buccal cells (Lucentini et al 2006 ), ray mucus (Kashiwagi et al 2015 ), lavral homogenate and milt (Navaneeth Krishnan et al 2016 ) and a big variety of tissue and swab samples (Table 1 ). Insect dried samples used for analysis included insects crash (Lall et al 2010 ; Dickey et al 2012 ) or suspension/homogenate (Snowden et al 2002 ; Harvey 2005 ; Desloire et al 2006 ), dry fecal spots (Russomando et al 1996 ; Machado et al 2000 ; Dorn et al 2001 ; Brito et al 2008 ; Braz et al 2008 ), haemolymph (Machado et al 2000 ), gut smear (Boid et al 1999 ; Adams et al 2006 ; Fall et al 2012 ), midgut and proboscises (Gillingwater et al 2010 ), crashed abdomen (Niare et al 2017 ), blood meal specimens (Reeves et al 2016 ) and saliva (Hall-Mendelin et al 2010 ; van den Hurk et al 2014 ; Flies et al 2015 ; Johnson et al 2015 ; Burkett-Cadena et al 2016 ; Kurucz et al…”