Comparison of PCR testing methods for white spot syndrome virus

In Thailand, several PCR-based methods are used by private and public service laboratories for the detection of white spot

syndrome virus (WSSV) infection in penaeid shrimp post larvae (PL) before they are stocked in rearing ponds. Conflicting test

results for similar samples sent to two service laboratories has decreased confidence in PCR testing. Thus, we compared the

sensitivity of several PCR methods commonly employed in Thailand using Taqman real-time PCR as the gold standard with a

purified WSSV template stock. Using this stock for assays, we found no significant inhibitory effect by WSSV-free host shrimp

DNA over the range 0 to 300 ng per reaction or by added DNA from WSSV-infected shrimp. Real-time PCR could detect WSSV

with certainty at dilutions of approximately 5 copies per reaction while 1000 copies were needed for a common one-step PCR

method and 50 for a common single-tube nested PCR (1N-PCR) method. Of 2 two-tube nested PCR protocols tested, one required

100 and the other 1000 copies. In addition to these sensitivity tests, a triple-blind ring test was carried out employing sets of 10

WSSV-infected DNA extracts sent to 12 commercial and public laboratories in Thailand, without specifying the PCR method to be

used. Returned results included no false positives and two false negatives, the latter both from light infection vials. This translated

into a test sensitivity of 97.3% and a specificity of 100%. Overall, the results confirmed the validity of PCR-based methods in

Thailand for detection of WSSV in shrimp DNA extracts.

© 2005 Elsevier B.V. All rights reserved.

Keywords: PCR-based methods; White spot syndrome virus (WSSV) infection; Penaeus monodo

3 responses to “Comparison of PCR testing methods for white spot syndrome virus

  1. T. Saiful Rizal

    Saya sedang tertarik mempelajari tentang Perikanan, baik budidaya serta saran dan prasarananya.
    Ada satu hal untuk saya ketahui dan mohon bantuan penjelasannya tentang “Shrimple” atau alat untuk test WSSV terhadap benur udang (EnBio Shimp Virus Detector Kit), untuk jenis alat tersebut Merek apa yang bagus/cocok digunakan dan cara penggunaannya. Alat tersebut dimana bisa kami dapatkan untuk kami beli.

    Atas penjelasan dan informasi saya ucapkan banyak terima kasih.

    T. Saiful Rizal
    081360243518/081377162740

    Dear all

    “Shrimple” merupakan salah satu alat deteksi untuk penyakit WSSV yang sangat cepat dalam waktu 5 menit anda sudah bisa mengetahui hasilnya…..alat ini cocok untuk aplikasi dilapangan. Alat ini bekerja berdasarkan sistem imunology atau anti body udang terhadap WSSV…….. dan sensitivitas alat ini sama dengan one step PCR.

    Jika jumlah copy virus yang menginfeksi udang banyak maka dapat terdeteksi tapi jika jumlah copy virusnya sedikit maka akan tidak terdeteksi…….

    produk dari en biotech sudah cukup bagus saya kira… bahkan pernah di masukan di juranal Aquacutture mengenai sensitivitasnya…..mungkin lain kali saya akan postingkan cara kerja SHRIMPLE beserta papernya

  2. T. Saiful Rizal

    Saya sedang tertarik mempelajari tentang Perikanan, baik budidaya serta saran dan prasarananya.
    Ada satu hal untuk saya ketahui dan mohon bantuan penjelasannya tentang “Shrimple” atau alat untuk test WSSV terhadap benur udang (EnBio Shimp Virus Detector Kit), untuk jenis alat tersebut Merek apa yang bagus/cocok digunakan dan cara penggunaannya.

  3. bang didik (pati)

    Reading the story in Thailand, what do you think about conflicting PCR which is occuring in Indonesian PCR Laboratory ? Have u ever try to reveal in which part the problem is apparently happen?? Coz i think it’s pathetic u know..

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