Journal of Arthropod-Borne Diseases 2011. 5(2):42-50.

Utility of Filter Paper for Preserving Insects, Bacteria, and Host Reservoir DNA for Molecular Testing
F Karimian, MM Sedaghat, MA Oshaghi, F Mohtarami, A Sanei Dehkordi, M Koosha, S Akbari, SS Hashemi-Aghdam

Abstract


Background: Appropriate methodology for storage biological materials, extraction of DNA, and proper DNA preservation is vital for studies involving genetic analysis of insects, bacteria, and reservoir hosts as well as for molecular diagnostics of pathogens carried by vectors and reservoirs. Here we tried to evaluate the utility of a simple filter paper-based for storage of insects, bacteria, rodent, and human DNAs using PCR assays.
Methods: Total body or haemolymph of individual mosquitoes, sand flies or cockroaches squashed or placed on the paper respectively. Extracted DNA of five different bacteria species as well as blood specimens of human and great gerbil Rhombomys opimus was pipetted directly onto filter paper. The papers were stored in room temperature up to 12 months during 2009 until 2011. At monthly intervals, PCR was conducted using a 1-mm disk from the DNA impregnated filter paper as target DNA. PCR amplification was performed against different target genes of the organisms including the ITS2-rDNA of mosquitoes, mtDNA-COI of the sand flies and cockroaches, 16SrRNA gene of the bacteria, and the mtDNA-CytB of the vertebrates.

Results: Successful PCR amplification was observed for all of the specimens regardless of the loci, taxon, or time of storage. The PCR amplification were ranged from 462 to 1500 bp and worked well for the specified target gene/s. Time of storage did not affect the amplification up to one year.
Conclusion: The filter paper method is a simple and economical way to store, to preserve, and to distribute DNA samples for PCR analysis.


Keywords


DNA, PCR, Insects, Bacteria, Vertebrate, Preservation, Filter paper,

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References


Adams ER, Malele II, Msangi AR, Gibson WC (2006) Trypanosome identifica- tion in wild tsetse populations in Tanzania using generic primers to am- plify the ribosomal RNA ITS-1 region. Acta Trop. 100: 103–109.

Ansell J, Hu JT , Gilbert SC, Hamilton KA, Hill AV, Lindsay SW (2000) Im- proved method for distinguishing the human source of mosquito blood meals between close family members. Trans Royal Soc Trop Med Hyg. 94:572–574.

Ataei S, Nateghpour M, Hajjaran H, Edris- sian G, Foroushani AR (2011) High specificity of semi-nested multiplex PCR using dried blood spots on DNA Banking Card in comparison with fro- zen liquid blood for detection of Plas- modium falciparum and Plasmodium vivax. J Clin Lab Anal. 25: 185–190.

Boakye DA, Tang J, Truc P, Merriweather A, Unnasch TR (1999) Identification of blood meals in haematophagous Diptera by Cytochrome b heteroduplex analysis. Med Vet Entomol. 13: 282–287.

Dobbs LJ, Madigan MN, Carter AB, Earls L (2002) Use of FTA gene guard filter paper for the storage and transporta- tion of tumor cells for molecular test- ing. Arch Path Lab Med. 126: 56–63.

Devost NC, Choy FY (2000) Mutation analysis of Gaucher disease using dot- blood samples on FTA filter paper. Am J Med Genet. 94: 417–420.

Favia G, Ricci I, Damiani C, Raddadi N, Crotti E, Marzorati M, Rizzi A, Urso R, Brusetti L, Borin S, Mora D, Scuppa P, Pasqualini L, Clementi E, Genchi M, Corona S, Negri I, Grandi G, Alma A, Kramer L, Esposito F, Bandi C, Sacchi L, Daffonchio D (2007) Bacteria of the genus Asaia stably associate with Ano- pheles stephensi, an Asian malarial mosquito vector. Proc Natl Acad Sci USA. 104: 9047–9051.

Fischer P, Wibowo H, Pischke S, Rückert P,Liebau E, Ismid IS, Supali T (2002) PCR-based detection and identification of the filarial parasite Brugia timori from Alor Island, Indonesia. Ann Trop Med Parasit. 96: 809–821.

Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cyto- chrome c oxidase subunit I from di- verse metazoan invertebrates. Mol Ma- rine Biol Biotech. 3: 294–299.

Gonzales JL, Loza A, Chacon E (2006) Sensitivity of different Trypanosoma vivax specific primers for the diagnosis of livestock trypanosomosis using dif- ferent DNA extraction methods. Vet Parasitol. 136: 119–126.

Harvey ML (2005) An alternative for the extraction and storage of DNA from insects in forensic entomology, J Fo- rensic Sci. 50: 627–629.

Hide G, Hughes JM, McNuff R (2003) A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper. BMC Ecol. 3: 7.

Lampel A, Orlandi PA, Kornegay L (2000) Improved template preparation for PCR based assays for detection of food bourne bacterial pathogens. App Envi- ron Microb. 66: 4539– 4542.

Lekweiry KM, Abdallahi MO, Ba H, Arnathau C, Durand P, Trape JF, Salem AO (2009) Preliminary study of malaria incidence in Nouakchott, Mau- ritania. Malaria J. 8: 92.

Litvaitis MK, Nunn G, Thomas WK, Kocher TD (1994) A molecular approach for the identification of meiofaunal tur- bellarians (Piatyhelminthes, Turbellaria). Marine Biol. 120: 437–442.

Maleki-Ravasan M, Oshaghi MA, Javadian E, Rassi Y, Sadraei J, Mohtarami F (2009) Blood Meal Identification in Field-Captured Sand flies: Comparison of PCR-RFLP and ELISA Assays. Iran J Arthropod-Borne Dis. 3: 8–18.

Mehravaran A, Oshaghi MA, Vatandoost H, Abai MR, Ebrahimzadeh A, Moazeni Roodi, A, Grouhi A (2011) First report on Anopheles fluviatilis U in south- eastern Iran. Acta Trop. 117: 76–81.

Natarajan P, Trinh T, Mertz L, Goldsbor- ough M, Fox DK (2000) Paper-based archiving of mammalian and plant samples for RNA analysis. Biotech- niques. 29: 1328–1333.

Nakazawa S, Marchand RP, Quang NT, Culleton R, Manh ND, Maeno Y (2009) Anopheles dirus co-infection with human and monkey malaria para- sites in Vietnam. Int J Parasitol. 39:1533–1537.

Orlandi PA, Lampel KA (2000) Extraction- free, filter-based template preparation

for rapid and sensitive PCR detection of pathogenic parasitic protozoa. J Clin Microbiol. 38: 2271–2277.

Oshaghi MA, Sedaghat MM, Vatandoost H (2003) Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran. East Mediterr Health J. 9: 659–666.

Oshaghi MA, Chavshin AR, Vatandoost H (2006) Analysis of mosquito blood meals using RFLP Markers. Exp Parasitol. 114: 259–264.

Oshaghi MA, Yaghobi-Ershadi MR, Shem- shad K, Pedram M, Amani H (2008) The Anopheles superpictus complex: introduction of a new malaria vector complex in Iran. Bull Soc Path Exo.101: 429–434.

Oshaghi MA, Maleki Ravasan N, Javadian EZ, Mohebali M, Hajjaran H, Zare Z, Mohtarami F, Rassi Y (2009) Vector incrimination of sand flies in the most important visceral Leishmaniasis focus in Iran. Am J Trop Med Hyg. 81: 572–577.

Oshaghi MA, Rasolian M, Shirzadi MR, Mohtarami F, Doosti S (2010) First report on isolation of Leishmania tropica from sandflies of a classical urban Cutaneous leishmaniasis focus in south- ern Iran. Exp Parasitol. 126: 445–450.

Oshaghi MA, Rassi Y, Tajedin L, Abai MR, Akhavan AA, Mohtarami F (2011a) Mitochondrial DNA diversity in the populations of great gerbils, Rhombo- mys opimus, main reservoir of cutane- ous leishmaniasis. Acta Trop. 119(2–3):165-17.

Oshaghi MA, Vatandoost H, Gorouhi A, Abai MR, Madjidpour A, Arshi S, Sadeghi H, Nazari M, Mehravaran A (2011b) Anopheline species composi- tion in border line of Iran-Azerbaijan. Acta Trop. 119(1): 44–49.

Rao RU, Atkinson LJ, Ramzy RM, Helmy H, Farid HA, Bockarie MJ, Susapu M,Laney SJ, Williams SA, Weil GJ (2006) A real-time PCR-based assay for detection of Wuchereria bancrofti DNA in blood and mosquitoes. Am J Trop Med Hyg. 74: 826–832.

Rogers CD, Burgoyne LA (1997) Bacterial typing: storing and processing of stabi- lized reference bacteria for polymerase chain reaction without preparing DNA- an example of an automatible proce- dure. Anal Biochem. 247: 223–227.

Rogers CD, Burgoyne LA (2000) Reverse transcription of an RNA genome from databasing paper (FTA). Biotechnol Appl Bioc. 31: 219–224.

Syafruddin D, Asih PB, Wahid I, Dewi RM, Tut,i S, Laowo I, Hulu W, Zendrato P, Laihad F, Shankar AH (2007) Malaria prevalence in Nias District, North Su- matra Province, Indonesia. Malaria J.6: 116.

Sant'Anna MR, Jones NG, Hindley JA, Mendes-Sousa AF, Dillon RJ, Cavalcante RR, Alexander B, Bates PA (2008) Blood meal identification and parasite detection in laboratory-fed and field- captured Lutzomyia longipalpis by PCR using FTA databasing paper. Acta Trop.107: 230–237.

Schaefer KU, Schoone GJ, Gachihi GS, Muller AS, Kager PA, Meredith SE (1995) Visceral leishmaniasis: use of the polymerase chain reaction in an epidemiological study in Baringo District, Kenya. T Roy Soc Trop Med H. 89: 492–495.

Smith LM, Burgoyne LA (2004) Collecting, archiving and processing DNA from wildlife samples using FTA databasing paper. BMC Ecol. 84: 4.

Tabachnick WJ (2010) Challenges in pre- dicting climate and environmental ef- fects on vector-borne disease episys- tems in a changing world. J Exp Biol.213: 946–954.

WHO (2009) World malaria report, WHO Press, World Health Organization, Geneva.

WHO (2000) The World Health Report,

WHO Press, World Health Organiza- tion, WHO Health Report- Prelims i- ixx/E.

Weigle KA, Labrada LA, Lozano K, Santrich C, Barker DC (2002) PCR Based Diagnosis of Acute and Chronic Cutaneous Leishmaniasis Caused by Leishmania (Viannia). J Clin Microbiol.40: 601–606.


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