Factors that can interfere with virus concentration from wastewater when using zeta plus 60S filter membranes

Queiroz, Ana Paula Spolidoro; Santos, Flávia Carvalho; Hársi, Charlotte Marianna; Candeias, J. A. N.; Monezi, Telma Alves; Mehnert, Dolores Ursula

doi:10.1590/s0074-02762000000500018

Cited by 3 publications

(2 citation statements)

References 7 publications

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“…The aqueous phase was transferred to another tube and treated as before. In method 2, microporous and positively charged membranes, Zeta Plus 60S, were rinsed with 10 ml of distilled water/cm 2 prior to use (24). Then, 50-ml autoclaved and nonautoclaved water samples were filtrated through a 65-cm 2 membrane.…”

Section: Methodsmentioning

confidence: 99%

“…Both studies reported a high rate of inhibition of RT-PCR after concentration of surface water samples on electropositive filters. Recently, our group observed that the electropositive filter membranes (ZP60S) release cytotoxic residues during the sample filtration procedure, although this problem was overcome by rinsing the membrane with distilled water before use, eliminating the cytotoxic substances from the membrane matrix (24). Probably the PCR inhibitors, such as humic substances and other organic and inorganic compounds (1,16), were eliminated, contributing to the good results obtained with our method.…”

Section: Figmentioning

confidence: 95%

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Electropositive Filter Membrane as an Alternative for the Elimination of PCR Inhibitors from Sewage and Water Samples

Queiroz

Santos

Sassaroli

et al. 2001

Appl Environ Microbiol

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PCR is the best method for the detection of enteric viruses present at low concentrations in environmental samples. However, some organic and inorganic compounds present in these samples can interfere in the reaction. Many of these substances are cytotoxic, too. The ZP60S filter membranes used in addition to fluorpentane treatment are quite efficient for virus concentration and simultaneous elimination of cytotoxicity from environmental samples. In this study, both procedures were used to promote the elimination of reverse transcriptase PCR (RT-PCR) inhibitors from sewage and sewage-polluted creek water. Samples were subjected separately to each of the following procedures: filtration through electropositive filter membranes (ZP60S), organic extraction with Vertrel XF, and filtration through ZP60S followed by organic extraction. Afterwards, aliquots were experimentally inoculated with rotavirus SA-11 RNA and subjected to RT-seminested PCR for amplification of the VP7 gene. Results showed that the ZP60S membranes efficiently eliminated the RT-PCR inhibitors from water samples. The sample processing method was also applied to 31 in natura sewage and creek water samples for detection of naturally occurring rotavirus. A duplex seminested PCR was used for the quick detection of couples of the four rotavirus genotypes (G1 to G4). Eight samples (25.8%) were positive, and rotavirus sequences were not detected in 23 (74.2%). Results were confirmed by direct immunoperoxidase method. In summary, the use of electropositive filter membrane is appropriate for the elimination of substances that can interfere with RT-PCR, obviating additional sample purification methods.

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Section: Methodsmentioning

confidence: 99%

Section: Figmentioning

confidence: 95%

Electropositive Filter Membrane as an Alternative for the Elimination of PCR Inhibitors from Sewage and Water Samples

Queiroz

Santos

Sassaroli

et al. 2001

Appl Environ Microbiol

View full text Add to dashboard Cite

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Enteroviruses in Water: Concentration and Detection

Goyal

2003

Encyclopedia of Environmental Microbiology

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Electronegative Filters Comparative Studies Electropositive Filters Other Media Factors Influencing Virus Concentration Second‐Step Concentration Molecular Methods of Virus Detection Inhibitors of RT‐PCR

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Detection ofThaumatotibia(Cryptophlebia)leucotretaGranulovirus DNA in Water Samples in an Insect Rearing Facility

Tobin¹,

Saayman

Boersma

et al. 2016

African Entomology

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Factors that can interfere with virus concentration from wastewater when using zeta plus 60S filter membranes

Abstract: Zeta plus filter membranes (ZP60S) have

Cited by 3 publications

References 7 publications

Electropositive Filter Membrane as an Alternative for the Elimination of PCR Inhibitors from Sewage and Water Samples

Electropositive Filter Membrane as an Alternative for the Elimination of PCR Inhibitors from Sewage and Water Samples

Enteroviruses in Water: Concentration and Detection

Detection ofThaumatotibia(Cryptophlebia)leucotretaGranulovirus DNA in Water Samples in an Insect Rearing Facility

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