Validation of a commercial insulated isothermal PCR-based POCKIT test for rapid and easy detection of white spot syndrome virus infection in Litopenaeus vannamei

Yun Long Tsai, Han Ching Wang, Chu Fang Lo, Feng-Jyu Tang-Nelson, Donald V Lightner, Bor Rung Ou, Ai Ling Hour, Chuan Fu Tsai, Cheng Chi Yen, Hsiao Fen Grace Chang, Ping Hua Teng, Pei Yu Lee

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Timely pond-side detection of white spot syndrome virus (WSSV) plays a critical role in the implementation of bio-security measures to help minimize economic losses caused by white spot syndrome disease, an important threat to shrimp aquaculture industry worldwide. A portable device, namely POCKIT™, became available recently to complete fluorescent probe-based insulated isothermal PCR (iiPCR), and automatic data detection and interpretation within one hour. Taking advantage of this platform, the IQ Plus™ WSSV Kit with POCKIT system was established to allow simple and easy WSSV detection for on-site users. The assay was first evaluated for its analytical sensitivity and specificity performance. The 95% limit of detection (LOD) of the assay was 17 copies of WSSV genomic DNA per reaction (95% confidence interval [CI], 13 to 24 copies per reaction). The established assay has detection sensitivity similar to that of OIE-registered IQ2000™ WSSV Detection and Protection System with serial dilutions of WSSV-positive Litopenaeus vannamei DNA. No cross-reaction signals were generated from infectious hypodermal and haematopoietic necrosis virus (IHHNV), monodon baculovirus (MBV), and hepatopancreatic parvovirus (HPV) positive samples. Accuracy analysis using700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5% (95% CI: 90.61-95.56%) and specificity 97% (95% CI: 94.31-98.50%). Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.

Original languageEnglish (US)
Article numbere90545
JournalPLoS One
Volume9
Issue number3
DOIs
StatePublished - Mar 13 2014

Fingerprint

White spot syndrome virus 1
White spot syndrome virus
Litopenaeus vannamei
Virus Diseases
Viruses
Polymerase Chain Reaction
infection
testing
Assays
portable equipment
confidence interval
assays
Confidence Intervals
Densovirinae
detection limit
analytical specificity
white spot syndrome
Security Measures
Monodon
shrimp fisheries

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Validation of a commercial insulated isothermal PCR-based POCKIT test for rapid and easy detection of white spot syndrome virus infection in Litopenaeus vannamei. / Tsai, Yun Long; Wang, Han Ching; Lo, Chu Fang; Tang-Nelson, Feng-Jyu; Lightner, Donald V; Ou, Bor Rung; Hour, Ai Ling; Tsai, Chuan Fu; Yen, Cheng Chi; Chang, Hsiao Fen Grace; Teng, Ping Hua; Lee, Pei Yu.

In: PLoS One, Vol. 9, No. 3, e90545, 13.03.2014.

Research output: Contribution to journalArticle

Tsai, Yun Long ; Wang, Han Ching ; Lo, Chu Fang ; Tang-Nelson, Feng-Jyu ; Lightner, Donald V ; Ou, Bor Rung ; Hour, Ai Ling ; Tsai, Chuan Fu ; Yen, Cheng Chi ; Chang, Hsiao Fen Grace ; Teng, Ping Hua ; Lee, Pei Yu. / Validation of a commercial insulated isothermal PCR-based POCKIT test for rapid and easy detection of white spot syndrome virus infection in Litopenaeus vannamei. In: PLoS One. 2014 ; Vol. 9, No. 3.
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abstract = "Timely pond-side detection of white spot syndrome virus (WSSV) plays a critical role in the implementation of bio-security measures to help minimize economic losses caused by white spot syndrome disease, an important threat to shrimp aquaculture industry worldwide. A portable device, namely POCKIT™, became available recently to complete fluorescent probe-based insulated isothermal PCR (iiPCR), and automatic data detection and interpretation within one hour. Taking advantage of this platform, the IQ Plus™ WSSV Kit with POCKIT system was established to allow simple and easy WSSV detection for on-site users. The assay was first evaluated for its analytical sensitivity and specificity performance. The 95{\%} limit of detection (LOD) of the assay was 17 copies of WSSV genomic DNA per reaction (95{\%} confidence interval [CI], 13 to 24 copies per reaction). The established assay has detection sensitivity similar to that of OIE-registered IQ2000™ WSSV Detection and Protection System with serial dilutions of WSSV-positive Litopenaeus vannamei DNA. No cross-reaction signals were generated from infectious hypodermal and haematopoietic necrosis virus (IHHNV), monodon baculovirus (MBV), and hepatopancreatic parvovirus (HPV) positive samples. Accuracy analysis using700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5{\%} (95{\%} CI: 90.61-95.56{\%}) and specificity 97{\%} (95{\%} CI: 94.31-98.50{\%}). Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.",
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AU - Lo, Chu Fang

AU - Tang-Nelson, Feng-Jyu

AU - Lightner, Donald V

AU - Ou, Bor Rung

AU - Hour, Ai Ling

AU - Tsai, Chuan Fu

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AU - Chang, Hsiao Fen Grace

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AU - Lee, Pei Yu

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