Parasitology Research

, Volume 102, Issue 3, pp 381–387 | Cite as

CP2 gene as a useful viability marker for Cryptosporidium parvum

  • Soo-Ung Lee
  • Migyo Joung
  • Myoung-Hee Ahn
  • Sun Huh
  • Hyunje Song
  • Woo-Yoon Park
  • Jae-Ran Yu
Original Paper

Abstract

The validity of the CP2 gene of Cryptosporidium parvum as a viability marker was evaluated using absolute quantitative real-time polymerase chain reaction (qPCR) assays. Total ribonucleic acid (RNA) was isolated from live and heat-killed C. parvum oocysts, and complementary deoxyribonucleic acid was synthesized and used as a template. The most accurate number of viable C. parvum oocysts was predicted when the CP2 gene was used as a target gene. The lower detection limit of the CP2 gene was ten oocysts, which was the most sensitive among examined target genes. With heat shock induction, only hsp70 messenger RNA (mRNA) was induced, and the predicted viable oocyst number was increased by heat shock for this marker. The CP2, hsp70, Cryptosporidium oocyst wall protein, and β-tubulin mRNAs were not detected in heat-killed oocysts, but the 18S ribosomal ribonucleic acid (rRNA) showed heat stability until 48 h after heat killing. Although the 18S rRNA demonstrated the fastest response in crossing point (CP) value among the examined primer sets in qPCR, overestimation of viable oocysts was noted in the analysis with this gene. In conclusion, the CP2 gene was identified as the most sensitive, reliable, and accurate candidate of a viability marker of C. parvum by qPCR evaluation.

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Soo-Ung Lee
    • 1
  • Migyo Joung
    • 1
  • Myoung-Hee Ahn
    • 2
  • Sun Huh
    • 3
  • Hyunje Song
    • 4
  • Woo-Yoon Park
    • 5
  • Jae-Ran Yu
    • 1
  1. 1.Department of Environmental and Tropical MedicineKonkuk University School of MedicineChungjuRepublic of Korea
  2. 2.Department of Parasitology, College of MedicineHanyang UniversitySeoulRepublic of Korea
  3. 3.Department of Parasitology, College of MedicineHallym UniversityChuncheonRepublic of Korea
  4. 4.Departmetnt of PathologyGwangju Health CollegeGwangjuRepublic of Korea
  5. 5.Department of Radiation Oncology, College of MedicineChungbuk National UniversityCheongjuKorea

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