Real-time PCR detection of adenoviruses, polyomaviruses, and torque teno viruses in river water in Japan

The aim of this study was to investigate the persistence of SARS-CoV-2 in post-mortem swabs of subjects who died from SARS-CoV-2 infection. The presence of the virus was evaluated post-mortem from airways of 27 SARS-CoV-2 positive patients at three different time points (T1 2 h; T2 12 h; T3 24 h) by real-time PCR. Detection of antibodies to SARS-CoV-2 was performed by Maglumi 2019-nCoV IgM/IgG chemiluminescence assay. SARS-CoV-2 viral RNA was still detectable in 70.3% of cases within 2 h after death and in 66,6% of cases up to 24 h after death. Our data showed an increase of the viral load in 78,6% of positive individuals 24 h post-mortem (T3) in comparison to that evaluated 2 h after death (T1). Noteworthy, we detected a positive T3 post-mortem swab (24 h after death) from 4 subjects who were negative at T1 (2 h after death). The results of our study may have an important value in the management of deceased subjects not only with a suspected or confirmed diagnosis of SARS-CoV-2, but also for unspecified causes and in the absence of clinical documentation or medical assistance.

Download Full-text

Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus

Molecular Plant Pathology ◽

10.1111/j.1364-3703.2007.00434.x ◽

2007 ◽

Vol 8 (6) ◽

pp. 803-809 ◽

Cited By ~ 42

Author(s):

CECILE FRANÇOIS ◽

CHANTAL CASTAGNONE ◽

NEIL BOONHAM ◽

JENNY TOMLINSON ◽

REBECCA LAWSON ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Satellite Dna ◽

Bursaphelenchus Xylophilus ◽

Pcr Detection ◽

Pinewood Nematode

Download Full-text

Real-time PCR detection of five prevalent bacteria causing acute meningitis

Apmis ◽

10.1111/j.1600-0463.2009.02539.x ◽

2009 ◽

Vol 117 (11) ◽

pp. 856-860 ◽

Cited By ~ 17

Author(s):

SARA THULIN HEDBERG ◽

PER OLCÉN ◽

HANS FREDLUND ◽

PAULA MÖLLING

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Detection ◽

Acute Meningitis

Download Full-text

Nanolitre real-time PCR detection of bacterial, parasitic, and viral agents from patients with diarrhoea in Nunavut, Canada

International Journal of Circumpolar Health ◽

10.3402/ijch.v72i0.19903 ◽

2013 ◽

Vol 72 (1) ◽

pp. 19903 ◽

Cited By ~ 20

Author(s):

David M. Goldfarb ◽

Brent Dixon ◽

Ioana Moldovan ◽

Nicholas Barrowman ◽

Kirsten Mattison ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Detection

Download Full-text

Rapid, specific and quantitative polymerase chain reaction (PCR) detection of pathogenic protozoa Entamoeba histolytica for drinking water supply

Water Science & Technology Water Supply ◽

10.2166/ws.2011.057 ◽

2011 ◽

Vol 11 (4) ◽

pp. 418-425 ◽

Cited By ~ 1

Author(s):

S. W. Lam ◽

H. B. Zhang ◽

L. Yu ◽

C. H. Woo ◽

K. N. Tiew ◽

...

Keyword(s):

Real Time ◽

Water Samples ◽

Real Time Pcr ◽

Genomic Dna ◽

Tap Water ◽

Pcr Detection ◽

Raw Water ◽

Conventional Pcr ◽

Polymerase Chain ◽

Species Specific

In this study, a quantitative species-specific polymerase chain reaction (PCR) method to rapidly detect E. histolytica in water is developed. First, the specificity of E. histolytica PCR detection was verified by using species-specific primers of 16S-like rRNA genes to clearly differentiate it from the closely related amoebae species E. dispar and E. moshkovskii. The sensitivity of this method was subsequently determined using purified E. histolytica genomic DNA and culture cells as PCR reaction templates. Results indicated that conventional PCR visualized on 1% agarose gel was able to detect as low as 0.02 pg genomic DNA and 5 cells, while real-time PCR could detect 0.01 pg genomic DNA and 2 cells of E. histolytica. The protocols for E. histolytica PCR detection in real water samples were then optimized by spiking E. histolytica cells into tap water and reservoir raw water samples. A two-round centrifugation treatment to concentrate amoeba cells directly as a PCR template was the most effective way to detect E. histolytica in spiked tap water samples, while DNA extraction after concentrating amoeba cells was required for spiked reservoir raw water samples. The detection limit of 50 E. histolytica cells in 100 ml tap water was achieved in 2 h from sample collection to real-time PCR data readout. With these established protocols, 78 tap water samples, 11 reservoir raw water samples and 4 feed water samples from Singapore water supply systems were analyzed by both conventional PCR and real-time PCR methods. No E. histolytica cell was detected in tested samples.

Download Full-text