Yang Wei, Hite Scott, Huang Yung T
Department of Pathology, University Hospitals of Cleveland, Case Western Reserve University, Cleveland, OH 44106, USA.
J Clin Virol. 2005 Oct;34(2):125-8. doi: 10.1016/j.jcv.2005.02.008.
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.
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.
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.
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.
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.
在采用针对即刻早期抗原（IEA）进行染色的空斑小室法出现之前，从临床标本中检测人巨细胞病毒（HCMV）是一个缓慢的过程。尽管培养法仍被认为不够灵敏，但除血液样本外广泛应用于其他样本检测。因此，需要一种提高培养法灵敏度的方法。
对Diagnostic Hybrids公司生产的CMV预处理培养基TurboTreat进行评估，观察其在Mv1Lu、R-Mix和MRC-5细胞上对HCMV检测灵敏度的提升效果。
在48孔板中培养Mv1Lu、R-Mix和MRC-5细胞单层，分别用TurboTreat处理过夜（o/n）、4小时或不处理，然后接种先前检测为阳性的HCMV标本。过夜培养后，固定细胞、染色并计数阳性细胞。
对于HCMV临床分离株，用CMV TurboTreat处理4小时后，Mv1Lu和R-Mix细胞的检测率提高了2至3倍，处理过夜后提高了4至6倍，MRC-5细胞的提高幅度较小。对于冷冻临床标本，Mv1Lu细胞过夜处理、4小时处理或未处理分别检测到23、21和15个阳性标本。R-Mix细胞分别检测到19、18和14个阳性标本，MRC-5细胞分别检测到16、15和15个阳性标本。无论是否处理，在Mv1Lu过夜处理未检测到阳性的情况下，其他细胞系均未检测到阳性。
在Mv1Lu细胞上用CMV TurboTreat过夜预处理是最佳处理条件，显著提高了HCMV的检测率。即使对Mv1Lu细胞进行4小时预处理，其检测率也显著高于未处理的Mv1Lu、R-Mix和MRC-5细胞。用CMV TurboTreat对Mv1Lu细胞预处理4小时或更长时间可提高HCMV快速检测的灵敏度。
