A comparison of filter capture and precultivation for quantitative detection of infectious hematopoietic necrosis virus (IHNV) by using polymerase chain reaction (PCR) combined with the most probable number (MPN) method

被引：0

作者：

Yoshinaka, T ^{[1
]}

Yoshimizu, M ^{[1
]}

Ezura, Y ^{[1
]}

机构：

[1] Hokkaido Univ, Fac Fisheries, Microbiol Lab, Hakodate, Hokkaido 041, Japan

来源：

JOURNAL OF AQUATIC ANIMAL HEALTH | 1999年 / 11卷 / 02期

关键词：

D O I：

10.1577/1548-8667(1999)011<0154:ACOFCA>2.0.CO;2

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

Infectious hematopoietic necrosis virus (IHNV) was collected from phosphate-buffered solutions and water that contained low numbers of the virus either by precultivation in cell cultures or capture by membrane filtration. The RNA from IHNV present in precultures or on membrane filters was extracted, and a 510-base-pair segment of the viral gene coding for the N-protein was amplified by reverse transcription polymerase chain reaction (RT-PCR). The number of virus particles present in the original sample was then calculated by using the most probable number (MPN) method. Low numbers of IHNV as preset in the original inoculum were accurately estimated by this PCR-MPN with precultivation. The PCR-MPN was less effective with the filter capture than the precultivation method because the quantity of IHNV detected was only 1/10 of that found in the original inoculum.

引用

页码：154 / 157

页数：4