RT-PCR and cell culture infectivity assay to detect enteroviruses during drinking water treatment processes.
In this study, 62 water samples were collected from two water treatment plants (WTPs) in Suez Canal cities (Port Said and Ismaillia) and one plant in Cairo (Giza WTP) in addition to the beginning of the two Nile river branches (Rosetta and Damietta). Viruses were concentrated by adsorption-elution ethod sing 142 mm-diameter nitrocellulose membrane of 0.45 microm pore size and eluted with 3% beef extract at pH 9.5. The concentrated samples were inoculated for 3 successive passages in three cell culture types (Vero, BGM and RD). Enterovirus RNAs in CPE-induced samples were extracted by guanidinium thiocyanate/ phenol/chloroform and heat shock methods and detected by RT-PCR and neutralization test. The results showed that eight samples [14.5% (8/62)] contained enteroviruses most of them were polioviruses [87.5% (7/8)] and coxsackievirus type B2 [12.5% (1/8)]. The three cell cultures were of the same sensitivity to detect the isolated viruses. Also, RT-PCR followed by neutralization assay facilitates and accelerate the results. The guanidinium thiocyanate extraction method was more sensitive than heat shock method. The results turned our attention to review our technology of water treatment and disinfection step in addition to the selection of suitable intake for the drinking water treatment plants.
Ali
M A
MA
Environmental Virology Lab., Department of Water Pollution Researches, National Research Centre, 12311 Dokki, Cairo, Egypt.
El-Esnawy
N A
NA
Shoaeb
A R
AR
Ibraheim
M
M
El-Hawaary
S E
SE
D003470:Culture Media; D004534:Egypt; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D006801:Humans; D009500:Neutralization Tests / Q000379:methods / Q000592:standards; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D045483:Rivers / Q000821:virology; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D010948:Viral Plaque Assay / Q000379:methods* / Q000592:standards; D014776:Virus Cultivation; D014871:Water Microbiology; D018508:Water Purification / Q000379:methods
D003470:Culture Media; D012367:RNA, Viral