Assessment of a new qPCR tool for the detection and identification of the root-knot nematode Meloidogyne enterolobii by an international test performance study

Küçük Resim Yok

Tarih

2016

Dergi Başlığı

Dergi ISSN

Cilt Başlığı

Yayıncı

SPRINGER

Erişim Hakkı

info:eu-repo/semantics/closedAccess

Özet

Rapid and reliable tools for detection and identification of plant parasitic nematodes are needed to prevent the introduction and spread of quarantine nematodes. A fast and simple DNA extraction method for target nematodes in nematode suspensions obtained from soil samples and a new quantitative real-time PCR method (qPCR) for the specific detection, identification and potential quantification of M. enterolobii were tested in an inter-laboratory comparison (ring test) to allow for a thorough evaluation of these molecular diagnostic tools. A test performance study involving seven laboratories was conducted to validate the developed protocols and to identify possible difficulties when implemented by diagnostic laboratories or national reference centers. Validation included test performance in terms of accuracy, analytical specificity, analytical sensitivity, repeatability, and reproducibility as defined by European Plant Protection Organization (EPPO) standard PM7/98. All positive and negative results for detection, identification and specificity were consistent between different laboratories despite different equipment used. Accuracy of real-time PCR was 100 % because test results and accepted reference values were in agreement. Analytical sensitivity results also matched between laboratories independent of the equipment used. The smallest amount of target DNA tested, two second-stage juveniles of M. enterolobii in a background of 500 non-target nematodes, was reliably detected by all labs. In addition, the repeatability and reproducibility of test results between laboratories was 100 %, even at the limit of detection. Thus, the inter-laboratory comparison showed the robustness of the developed methods and confirmed the in-house validation data.

Açıklama

Anahtar Kelimeler

Real-time PCR, Performance assessment, Meloidogyne enterolobii, Sensitivity, Specificity

Kaynak

EUROPEAN JOURNAL OF PLANT PATHOLOGY

WoS Q Değeri

Q2

Scopus Q Değeri

Q1

Cilt

144

Sayı

1

Künye