TY - JOUR
T1 - Development of a highly sensitive method for the detection of cryptosporidium parvum virus type 1 (Cspv1)
AU - Shehata, Ayman Ahmed
AU - Bando, Hironori
AU - Fukuda, Yasuhiro
AU - Kabir, Mohammad Hazzaz Bin
AU - Murakoshi, Fumi
AU - Itoh, Megumi
AU - Fujikura, Atsushi
AU - Okawa, Hiroaki
AU - Endo, Takuto
AU - Goto, Akira
AU - Kachi, Masayuki
AU - Nakayama, Toshie
AU - Kano, Yuto
AU - Oishi, Shoko
AU - Otomaru, Konosuke
AU - Kazama, Kei
AU - Essa, Mohamed Ibrahim
AU - Kato, Kentaro
N1 - Funding Information:
The authors would like to thank the veterinarians of the different prefectures for collecting the specimens and collaborating with us. This study was funded by grants-in-aid for Scientific Research (B:17H03913) and (C:16KT0141) and Scientific Research on Innovative Areas (3805) from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan, and by a Livestock Promotional Subsidy from the Japan Racing Association. We would like to thank the Ministry of higher Education and Scientific Research, the Arab Republic of Egypt for support during this study in accordance with the Egypt-Japan Education Partnership (EJEP).
Publisher Copyright:
© 2020, Hokkaido University. All rights reserved.
PY - 2020
Y1 - 2020
N2 - Cryptosporidium is an apicomplexan zoonotic parasite that infects most mammals, including humans. Cryptosporidium parvum virus type 1 (CSpV1) is the first member within the Partitiviridae family recognized to infect protozoan hosts. Cryptosporidium tracking based on CSpV1 detection has been attempted; however, each study used different conditions for the PCR protocol, primers, and target viral sequences. Accordingly, the sensitivity of PCR-based CSpV1 detection remains unclear. In addition, oocyst purification from clinical samples can be problematic due to small number of oocysts, sample degradation and low yield efficiency of currently used purification methods. Here we show that the second half of the coding region of dsRNA2 can be detected from various types of clinical samples, without the need for oocyst purification, by using a semi-nested-PCR technique. Furthermore, we show that the short sequence targeted in this study has higher diversity than the Cryptosporidium GP60 gene. Taken together, our findings suggest that this method could be used as an important tracking marker for Cryptosporidium species.
AB - Cryptosporidium is an apicomplexan zoonotic parasite that infects most mammals, including humans. Cryptosporidium parvum virus type 1 (CSpV1) is the first member within the Partitiviridae family recognized to infect protozoan hosts. Cryptosporidium tracking based on CSpV1 detection has been attempted; however, each study used different conditions for the PCR protocol, primers, and target viral sequences. Accordingly, the sensitivity of PCR-based CSpV1 detection remains unclear. In addition, oocyst purification from clinical samples can be problematic due to small number of oocysts, sample degradation and low yield efficiency of currently used purification methods. Here we show that the second half of the coding region of dsRNA2 can be detected from various types of clinical samples, without the need for oocyst purification, by using a semi-nested-PCR technique. Furthermore, we show that the short sequence targeted in this study has higher diversity than the Cryptosporidium GP60 gene. Taken together, our findings suggest that this method could be used as an important tracking marker for Cryptosporidium species.
KW - CSpV1
KW - Calves
KW - Cryptosporidium
KW - PCR
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U2 - 10.14943/jjvr.68.3.159
DO - 10.14943/jjvr.68.3.159
M3 - Article
AN - SCOPUS:85092018310
VL - 68
SP - 159
EP - 170
JO - Japanese Journal of Veterinary Research
JF - Japanese Journal of Veterinary Research
SN - 0047-1917
IS - 3
ER -