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A simple, effective and inexpensive method to isolate the nucleic acid (DNA/RNA) from a single tick for molecular detection of various pathogens

Yıl 2024, Cilt: 10 Sayı: 1 - January 2024, 1 - 7, 04.01.2024
https://doi.org/10.18621/eurj.1315058

Öz

Objectives: Ticks are vectors of a number of pathogens, which cause diseases with fatal consequences, therefore it is essential to detect such pathogens from single tick. Molecular methods like polymerase chain reaction (PCR) are offering such possibilities. At present, cumbersome methods involving liquid nitrogen, cutting ticks with scalpels as well as pooled ticks are being used worldwide. Our goal is to develop a reliable and fast method to obtain nucleic acid (DNA/RNA) from the single tick shipped at room temperature to detect various pathogens.

Methods: We developed a mechanical crushing method with mini column nucleic acid isolation from a single tick shipped at room temperature with postal or currier service in a letter. PCR detection was done for Borrelia burgdorferi and tick-borne encephalitis virus as examples.

Results: This method was used successfully for the isolation of nucleic acid from single tick and later used for PCR detection of B. burgdorferi and tick-borne encephalitis virus on 17 single tick samples as examples, but for last 18 years, this method was used on more than 250 ticks from Germany. Spectrometric values indicate the presence of sufficient yield of DNA and RNA (up to 900 µg/mL per tick) during the isolation.

Conclusions: This may be the first report about a number of one single tick cases, which were sent at room temperature in letters with postal services for isolation of the nucleic acid with mini column kit and used later on for PCR detection of various pathogens. This inexpensive and simple method may be used in any laboratory worldwide for monitoring the presence of tick-borne pathogens.

Kaynakça

  • 1. Lee KO, Park MY, Kim LH, Seong HS, Park BH, Jeong SJ. Pandemic novel influenza A (H1N1) virus in Korea: the experience from August to September 2009. Korean J Clin Lab Sci. 2009;41(4):145-152.
  • 2. Muller MG, George AR, Walochnik J. Acinetobacter baumannii in localised cutaneous mycobacteriosis in falcons. Vet Med Int. 2010;2010:321797. doi: 10.4061/2010/321797.
  • 3. Marushchak L, Golovko A, Deriabin O. Epizootic situation and development tools for detection of Q-fever in Ukraine: Abstract number 21.049, International Meeting on Emerging Diseases and Surveillance: Vienna, Austria, February 15-18, 2013.
  • 4. Othman AA, Eissa AA, Markous RD, Ahmed BD, Al-Allawi NA. Hepatitis C virus genotypes among multiply transfused hemoglobinopathy patients from Northern Iraq. Asian J Transfus Sci. 2014;8(1):32-34. doi: 10.4103/0973-6247.126687.
  • 5. Colín-Ferreyra Mdel C, Mendieta-Zerón H, Romero-Figueroa Mdel S, Martínez-Madrigal M, Martínez-Pérez S, Domínguez-García MV. [Expression of gamma interferon during HPV and Chlamydia trachomatis infection in cervical samples]. Enferm Infecc Microbiol Clin. 2015;33(2):105-109. doi: 10.1016/j.eimc.2014.05.014. [Article in Spanish]
  • 6. Norouzian H, Ghobadian Diali H, Azadpour M, et al. PCR detection of Coxiella burnetii in milk samples of ruminants, Iran. J Med Bacteriol. 2018;7(1, 2):31-35.
  • 7. Reifenberger GC, Thomas BA, Rhodes DVL. Comparison of DNA extraction and amplification techniques for use with engorged hard-bodied ticks. Microorganisms. 2022;10(6):1254. doi: 10.3390/microorganisms10061254.
  • 8. Bhatia S. Pitfalls found in SARS CoV-2 specific test performance during the comparison between WHO recommended method and a commercial test. Atlantic J Med Sci Res 2023;3(1):22-26. doi: 10.5455/atjmed.2022.12.024
  • 9. Laroche M, Weeks, ENI. Vector-borne bacterial diseases: a neglected field of infectious diseases research. Med Vet Entomol. 2023;37(2):177-178. doi: 10.1111/mve.12646.
  • 10. Harvey E, Rose K, Eden J-S, et al. Extensive diversity of RNA viruses in Australian ticks. J Virol. 2019;93(3):e01358-18. doi: 10.1128/JVI.01358-18.
  • 11. Hubbard MJ, Cann, KJ, Wright DJM. Validation and rapid extraction of nucleic acids from alcohol-preserved ticks. Exp Appl Acarol. 1995;19(8):473-478. doi: 10.1007/BF00048266.
  • 12. Stanley H, Rhodes DVL. Presence of Rickettsia species in ticks collected from companion animals in Northeastern Georgia, United States. Vet Sci. 2021;8(3):37. doi: 10.3390/vetsci8030037.
  • 13. Cafiso A, Chiappa G, Luzzago C, et al. Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens. MethodsX. 2021;8:101315. doi: 10.1016/j.mex.2021.101315.
  • 14. Crowder CD, Rounds MA, Phillipson CA, et al. Extraction of total nucleic acids from ticks for the detection of bacterial and viral pathogens. J Med Entomol. 2010;47(1):89-94. doi: 10.1603/033.047.0112.
  • 15. Shan J, Jia Y, Hickenbotham P, Teulières L, Clokie MRJ. Combining citizen science and molecular diagnostic methods to investigate the prevalence of Borrelia burgdorferi s.l. and Borrelia miyamotoi in tick pools across Great Britain. Front Microbiol. 2023;14:1126498. doi: 10.3389/fmicb.2023.1126498.
  • 16. Jones AM, Van de Wyngaerde MT, Machtinger ET, Rajotte EG, Thomas C Baker TC. Choice of laboratory tissue homogenizers matters when recovering nucleic acid from medically important ticks. J Med Entomol. 2020;57(4):1221-1227. doi: 10.1093/jme/tjaa006.
  • 17. Day CA, Butler RA, Durick HE, et al. An ecological and epidemiological single-season survey of Anaplasma and Ehrlichia positive ticks in Victoria Falls National Park, Zimbabwe. Med Vet Entomol. 2023;37(2):195-208. doi: 10.1111/mve.12640.
  • 18. Ammazzalorso AD, Zolnik CP, Daniels TJ, Kolokotronis S. To beat or not to beat a tick: comparison of DNA extraction methods for ticks (Ixodes scapularis). PeerJ. 2015;3:e1147. doi: 10.7717/peerj.1147.
Yıl 2024, Cilt: 10 Sayı: 1 - January 2024, 1 - 7, 04.01.2024
https://doi.org/10.18621/eurj.1315058

Öz

Kaynakça

  • 1. Lee KO, Park MY, Kim LH, Seong HS, Park BH, Jeong SJ. Pandemic novel influenza A (H1N1) virus in Korea: the experience from August to September 2009. Korean J Clin Lab Sci. 2009;41(4):145-152.
  • 2. Muller MG, George AR, Walochnik J. Acinetobacter baumannii in localised cutaneous mycobacteriosis in falcons. Vet Med Int. 2010;2010:321797. doi: 10.4061/2010/321797.
  • 3. Marushchak L, Golovko A, Deriabin O. Epizootic situation and development tools for detection of Q-fever in Ukraine: Abstract number 21.049, International Meeting on Emerging Diseases and Surveillance: Vienna, Austria, February 15-18, 2013.
  • 4. Othman AA, Eissa AA, Markous RD, Ahmed BD, Al-Allawi NA. Hepatitis C virus genotypes among multiply transfused hemoglobinopathy patients from Northern Iraq. Asian J Transfus Sci. 2014;8(1):32-34. doi: 10.4103/0973-6247.126687.
  • 5. Colín-Ferreyra Mdel C, Mendieta-Zerón H, Romero-Figueroa Mdel S, Martínez-Madrigal M, Martínez-Pérez S, Domínguez-García MV. [Expression of gamma interferon during HPV and Chlamydia trachomatis infection in cervical samples]. Enferm Infecc Microbiol Clin. 2015;33(2):105-109. doi: 10.1016/j.eimc.2014.05.014. [Article in Spanish]
  • 6. Norouzian H, Ghobadian Diali H, Azadpour M, et al. PCR detection of Coxiella burnetii in milk samples of ruminants, Iran. J Med Bacteriol. 2018;7(1, 2):31-35.
  • 7. Reifenberger GC, Thomas BA, Rhodes DVL. Comparison of DNA extraction and amplification techniques for use with engorged hard-bodied ticks. Microorganisms. 2022;10(6):1254. doi: 10.3390/microorganisms10061254.
  • 8. Bhatia S. Pitfalls found in SARS CoV-2 specific test performance during the comparison between WHO recommended method and a commercial test. Atlantic J Med Sci Res 2023;3(1):22-26. doi: 10.5455/atjmed.2022.12.024
  • 9. Laroche M, Weeks, ENI. Vector-borne bacterial diseases: a neglected field of infectious diseases research. Med Vet Entomol. 2023;37(2):177-178. doi: 10.1111/mve.12646.
  • 10. Harvey E, Rose K, Eden J-S, et al. Extensive diversity of RNA viruses in Australian ticks. J Virol. 2019;93(3):e01358-18. doi: 10.1128/JVI.01358-18.
  • 11. Hubbard MJ, Cann, KJ, Wright DJM. Validation and rapid extraction of nucleic acids from alcohol-preserved ticks. Exp Appl Acarol. 1995;19(8):473-478. doi: 10.1007/BF00048266.
  • 12. Stanley H, Rhodes DVL. Presence of Rickettsia species in ticks collected from companion animals in Northeastern Georgia, United States. Vet Sci. 2021;8(3):37. doi: 10.3390/vetsci8030037.
  • 13. Cafiso A, Chiappa G, Luzzago C, et al. Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens. MethodsX. 2021;8:101315. doi: 10.1016/j.mex.2021.101315.
  • 14. Crowder CD, Rounds MA, Phillipson CA, et al. Extraction of total nucleic acids from ticks for the detection of bacterial and viral pathogens. J Med Entomol. 2010;47(1):89-94. doi: 10.1603/033.047.0112.
  • 15. Shan J, Jia Y, Hickenbotham P, Teulières L, Clokie MRJ. Combining citizen science and molecular diagnostic methods to investigate the prevalence of Borrelia burgdorferi s.l. and Borrelia miyamotoi in tick pools across Great Britain. Front Microbiol. 2023;14:1126498. doi: 10.3389/fmicb.2023.1126498.
  • 16. Jones AM, Van de Wyngaerde MT, Machtinger ET, Rajotte EG, Thomas C Baker TC. Choice of laboratory tissue homogenizers matters when recovering nucleic acid from medically important ticks. J Med Entomol. 2020;57(4):1221-1227. doi: 10.1093/jme/tjaa006.
  • 17. Day CA, Butler RA, Durick HE, et al. An ecological and epidemiological single-season survey of Anaplasma and Ehrlichia positive ticks in Victoria Falls National Park, Zimbabwe. Med Vet Entomol. 2023;37(2):195-208. doi: 10.1111/mve.12640.
  • 18. Ammazzalorso AD, Zolnik CP, Daniels TJ, Kolokotronis S. To beat or not to beat a tick: comparison of DNA extraction methods for ticks (Ixodes scapularis). PeerJ. 2015;3:e1147. doi: 10.7717/peerj.1147.

Ayrıntılar

Birincil Dil İngilizce
Konular Parazitoloji, Bulaşıcı Hastalıklar
Bölüm Original Article
Yazarlar

Sudhir BHATİA 0000-0002-9881-7101

Gudrun BAERSCH Bu kişi benim 0009-0007-1290-6458

Erken Görünüm Tarihi 11 Eylül 2023
Yayımlanma Tarihi 4 Ocak 2024
Gönderilme Tarihi 15 Haziran 2023
Kabul Tarihi 7 Eylül 2023
Yayımlandığı Sayı Yıl 2024 Cilt: 10 Sayı: 1 - January 2024

Kaynak Göster

AMA BHATİA S, BAERSCH G. A simple, effective and inexpensive method to isolate the nucleic acid (DNA/RNA) from a single tick for molecular detection of various pathogens. Eur Res J. Ocak 2024;10(1):1-7. doi:10.18621/eurj.1315058

e-ISSN: 2149-3189 


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