Enhancement of cytomegalovirus detection in mink lung cells using CMV Turbo.
BACKGROUND
Detection of HCMV from clinical specimens was a slow process until the development of shell vial method with staining for immediate early antigen (IEA). Culture though still considered insensitive is widely used for other than blood samples. A method to improve culture sensitivity is desirable.


OBJECTIVE
TurboTreat, a CMV pretreatment medium from Diagnostic Hybrids Inc., was evaluated on Mv1Lu, R-Mix and MRC-5 cells for improved sensitivity for HCMV detection.


METHODS
Monolayers of Mv1Lu, R-Mix and MRC-5 cells in 48-well plates were treated with TurboTreat for overnight (o/n), 4 h or left untreated and then inoculated with previously positive HCMV specimens. After o/n incubation, cells were fixed, stained and positive cells counted.


RESULTS
CMV TurboTreat enhanced detection 2-3-fold after 4 h treatment and 4-6-fold after o/n treatment in Mv1Lu and R-Mix cells and to a lesser extend in MRC-5 cells with clinical isolates of HCMV. With frozen clinical specimens, Mv1Lu cells treated o/n, 4 h or untreated detected 23, 21 and 15 positive specimens, respectively. R-Mix cells detected 19, 18, and 14 positives, respectively and MRC-5 cells detected 16, 15 and 15 positives, respectively. In no case was a positive detected in another cell line regardless of treatment that was not detected in Mv1Lu treated o/n.


CONCLUSIONS
The o/n pretreatment with CMV TurboTreat on Mv1Lu cells is the optimum condition of treatment and significantly enhanced the detection of HCMV. Even 4 h pretreatment of Mv1Lu cells showed significant enhancement over untreated Mv1Lu, R-Mix and MRC-5 cells. Pretreatment of Mv1Lu cells with CMV TurboTreat for 4 h or longer increased the sensitivity of rapid HCMV detection.
Yang
Wei
W
Department of Pathology, University Hospitals of Cleveland, Case Western Reserve University, Cleveland, OH 44106, USA.
Hite
Scott
S
Huang
Yung T
YT
D000818:Animals; D002460:Cell Line; D003470:Culture Media; D003587:Cytomegalovirus / Q000254:growth & development* / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology*; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D008907:Mink; D012680:Sensitivity and Specificity; D014776:Virus Cultivation / Q000379:methods*
D003470:Culture Media