Iranian Journal of Fisheries Sciences
مجله علوم شیلاتی ایران
IJFS
Agriculture
http://jifro.ir
1
admin
1562-2916
2322-5696
10.18869/acadpub.ijfs
en
jalali
1401
10
1
gregorian
2023
1
1
22
1
online
1
fulltext
en
Research Article: Establishment of pre-enrichment step and DNA extraction method for the novel PCR procedure of acute hepatopancreatic necrosis disease (AHPND) and mutant-AHPND fast detection in shrimp
Biotechnology
Biotechnology
پژوهشي
Orginal research papers
<span style="font-size:11pt"><span style="line-height:107%"><span style="font-family:Calibri,sans-serif"><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">After the first outbreak</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> of Acute Hepatopancreatic Necrosis Disease (AHPND) i</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">n China in 2009, </span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">this disease </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">has been still considered a globally dangerous disease in shrimp</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> aquaculture industry</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">. Currently, there is no effective method to prevent and treat AHPND. Therefore,</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> rapid</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> detection methods which could avert and control this disease in shrimp were considered the most effective strategy. In </span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">2021</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">, a new PCR reaction was established that can simultaneously detect AHPND and mutant-AHPND. Intending to develop a PCR kit, the PCR procedure including a pre-enrichment step and DNA</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">-</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">extract</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">ed</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> method</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> was </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">establish</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">ed</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> to conduct the PCR reaction. The new PCR procedure was verified with a detection limit of 5.10<sup>3</sup> CFU/mL</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">. </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">Th</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">is detection</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> limit was two times higher than the conventional PCR methods</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">,</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> which are currently used to detect AHPND. <i>V</i></span></span></span><i><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">ibrio</span></span></span></i><i><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> parahaemolyticus</span></span></span></i><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> showed the best growth at 3</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">7°C </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">in the broth with</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> hepatopancreas of</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> shrimp. A simple boiling method to extract DNA in bacterial broth with shrimp tissues was also</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"> modified</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">. The PCR procedure has successfully been validated on 42 AHPND-suspecting samples. These results would be developed into a novel standard procedure using a PCR kit for rapid detection of AHPND and </span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">related </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">mutant-AHPND for </span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">rapid </span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">disease diagnosis in shrimp farm</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">s</span></span></span><span lang="TR" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif">.</span></span></span> <span style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"Times New Roman",serif"></span></span></span></span></span></span>
AHPND, mutant-AHPND, DNA extraction, PCR, Vibrio parahaemolyticus
99
113
http://jifro.ir/browse.php?a_code=A-10-4238-1&slc_lang=en&sid=1
T.D.
Mai-Hoang
18c64007@student.hcmus.edu.vn
100319475328460046661
100319475328460046661
No
University of Science, VNU-HCM
M.H.T.
Nguyen
1718178@student.hcmus.edu.vn
100319475328460046662
100319475328460046662
No
University of Science, VNU-HCM
T.B.
Huynh
1718144@student.hcmus.edu.vn
100319475328460046663
100319475328460046663
No
University of Science, VNU-HCM
T.L.
Tran
tlthuoc@hcmus.edu.vn
100319475328460046664
100319475328460046664
No
University of Science, VNU-HCM
H.
Tran-Van
tvhieu@hcmus.edu.vn
100319475328460046665
100319475328460046665
Yes
University of Science, VNU-HCM