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用于从临床标本中检测和分型单纯疱疹病毒的ELVIS HSV ID/分型系统的评估

Evaluation of ELVIS HSV ID/Typing System for the detection and typing of herpes simplex virus from clinical specimens.

作者信息

Turchek B M, Huang Y T

机构信息

Department of Pathology, University Hospitals of Cleveland, Case Western Reserve University, OH 44106, USA.

出版信息

J Clin Virol. 1999 Jan;12(1):65-9. doi: 10.1016/s0928-0197(98)00066-x.

Abstract

BACKGROUND

Herpes simplex virus (HSV) is a common pathogen with two serotypes: HSV-1 and HSV-2. HSV infection does not pose much of a threat to an immunocompetent host but to an immunocompromised host or a neonate the infection can be fatal. The Enzyme-Linked Virus Inducible System (ELVIS) employs a genetically altered baby hamster kidney (BHK) cell line that allows for the rapid overnight detection of HSV but also includes an immunofluorescent stain for the simultaneous detection and typing of HSV-1 and HSV-2.

OBJECTIVE

To evaluate the ELVIS HSV ID/Typing System in comparison with HSV identification and typing in primary rabbit kidney (PRK) cells grown in shell vials.

STUDY DESIGN

Over a period of 6 weeks, 130 specimens were submitted to the diagnostic virology laboratory and cultured for the presence of HSV. Two PRK shell vials and one ELVIS BHK shell vial were inoculated with patient specimen. PRK shell vials were observed for cytopathic effect (CPE) for up to 4 days. When CPE was observed the PRK shell vials were fixed and one shell vial was stained with HSV-1 monoclonal antibody (Mab) and the other was stained with HSV-2 Mab. The coverslips were observed under the fluorescent microscope for specific apple-green fluorescence. The BHK shell vials were incubated overnight, fixed, and stained with galactopyranoside (X-Gal). If blue cells were present, the specimen was positive for HSV. The coverslip was then observed under the fluorescent microscope for the presence of specific apple-green fluorescence, indicating HSV-2. If no specific apple-green stain was observed, the coverslip was stained with a fluorescent conjugated goat anti-mouse IgG to determine the presence of HSV-1.

RESULTS

Of the 130 specimens, PRK shell vials detected 43 positive HSV; 30 were HSV-2 and 13 were HSV-1. The ELVIS BHK shell vials detected 42 positive HSV; 30 were HSV-2 and 12 were HSV-1. One low titer specimen was not identified as being HSV positive. Two specimens were not directly typed by the ELVIS system. One specimen had only one blue cell present and did not show specific staining for either HSV-1 or HSV-2. The other specimen had only five blue cells present and only one fluorescent cell present that was difficult to type. As suggested by the manufacturer's instructions, both specimens that were not directly typed were re-grown overnight from their supernatants and were correctly identified and typed.

CONCLUSION

The ELVIS HSV ID/Typing System is a rapid, highly specific and sensitive method of overnight HSV detection and typing.

摘要

背景

单纯疱疹病毒(HSV)是一种常见病原体,有两种血清型:HSV-1和HSV-2。HSV感染对免疫功能正常的宿主威胁不大,但对免疫功能低下的宿主或新生儿,感染可能是致命的。酶联病毒诱导系统(ELVIS)采用基因改造的幼仓鼠肾(BHK)细胞系,可在一夜之间快速检测HSV,还包括一种免疫荧光染色剂,用于同时检测和分型HSV-1和HSV-2。

目的

与在塑料培养瓶中生长的原代兔肾(PRK)细胞中进行HSV鉴定和分型相比,评估ELVIS HSV鉴定/分型系统。

研究设计

在6周的时间里,将130份标本提交给诊断病毒学实验室,培养以检测HSV的存在。将两份PRK塑料培养瓶和一份ELVIS BHK塑料培养瓶接种患者标本。观察PRK塑料培养瓶长达4天的细胞病变效应(CPE)。当观察到CPE时,将PRK塑料培养瓶固定,一份塑料培养瓶用HSV-1单克隆抗体(Mab)染色,另一份用HSV-2 Mab染色。在荧光显微镜下观察盖玻片,以观察特异性苹果绿荧光。将BHK塑料培养瓶孵育过夜,固定,并用吡喃半乳糖苷(X-Gal)染色。如果存在蓝色细胞,则标本HSV呈阳性。然后在荧光显微镜下观察盖玻片,以观察是否存在特异性苹果绿荧光,表明为HSV-2。如果未观察到特异性苹果绿染色,则用荧光偶联山羊抗小鼠IgG对盖玻片进行染色,以确定HSV-1的存在。

结果

在130份标本中,PRK塑料培养瓶检测到43份HSV阳性;30份为HSV-2,13份为HSV-1。ELVIS BHK塑料培养瓶检测到42份HSV阳性;30份为HSV-2,12份为HSV-1。一份低滴度标本未被鉴定为HSV阳性。两份标本未被ELVIS系统直接分型。一份标本仅存在一个蓝色细胞,未显示HSV-1或HSV-2的特异性染色。另一份标本仅存在五个蓝色细胞,仅存在一个难以分型的荧光细胞。按照制造商的说明,两份未直接分型的标本均从其上清液中重新培养过夜,并被正确鉴定和分型。

结论

ELVIS HSV鉴定/分型系统是一种快速、高度特异且敏感的过夜HSV检测和分型方法。

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