Detection of Herpes simplex virus (HSV) genome using polymerase chain reaction (PCR) in clinical samples Comparison of PCR with standard laboratory methods for the detection of HSV

Hn, Madhavan; Kannian, Priya; Anand, Appakkudal R.; Therese, K Lily

doi:10.1016/s1386-6532(99)00047-5

Cited by 53 publications

(28 citation statements)

References 14 publications

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“…The PCR assay amplifies a 179-bp region from the HSV DNA polymerase gene (5,10,12). Samples were extracted as described above.…”

Section: Specimensmentioning

confidence: 99%

“…Besides the potential for sensitivity roughly equivalent to gel detection, the use of target-specific probes in real-time PCR offers an additional level of specificity. These probes also allow the detection of single nucleotide polymorphisms (SNPs) using melt curve analysis, permitting typing of different viral species.While real-time PCR has been widely adopted for molecular infectious disease testing, real-time technology has often been compared to traditional methods such as culture, with a predictable enhancement in sensitivity (4,6,7,12). The current literature is more limited on the comparative performance of real-time PCR with established PCR formats.…”

mentioning

confidence: 99%

“…While real-time PCR has been widely adopted for molecular infectious disease testing, real-time technology has often been compared to traditional methods such as culture, with a predictable enhancement in sensitivity (4,6,7,12). The current literature is more limited on the comparative performance of real-time PCR with established PCR formats.…”

mentioning

confidence: 99%

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Effect of Sequence Polymorphisms on Performance of Two Real-Time PCR Assays for Detection of Herpes Simplex Virus

Stevenson¹,

Hymas²,

Hillyard

2005

J Clin Microbiol

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Herpes simplex virus (HSV) is the most common cause of acquired, sporadic encephalitis in the UnitedStates. PCR identification of HSV in spinal fluid has become the diagnostic gold standard due to its sensitivity and potential for speed, replacing other methods such as culture. We developed a real-time PCR assay to detect HSV, using a new type of hybridization probe, the Eclipse probe. In this study, we ran 323 samples (171 positives and 152 negatives) with the Eclipse real-time PCR assay and compared these results with another PCR assay using gel detection. The real-time assay agreed with our reference method for 319 out of the 323 samples tested (99%). Using two different real-time PCR platforms, we discovered that SNPs within the amplicon's probe binding region that are used to distinguish HSV-1 from HSV-2 can decrease assay sensitivity. This problem is potentially a general one for assays using fluorescent probes to detect target amplification in a real-time format. While real-time PCR can be a powerful tool in the field of infectious disease, careful sequence evaluation and clinical validation are essential in creating an effective assay.PCR is now accepted as the gold standard for diagnosis of herpes simplex virus (HSV) infections in the central nervous system (CNS), exhibiting a high degree of specificity and a sensitivity superior to that of culture or DFA (3,7,10,11,17,18). While viral culture for HSV encephalitis is still routinely employed in the clinical lab, it is only capable of detecting virus in less than 5% of patients with CNS infections (10, 14). Detection of low viral titers often found in spinal fluid requires the high level of sensitivity that is provided by amplificationbased assays (13).In recent years, real-time PCR has made testing more convenient. Real-time PCR is more amenable to automation, allowing higher throughputs and decreased turnaround times. In addition, the implementation of a one-tube closed reaction greatly reduces the possibility for contamination. Besides the potential for sensitivity roughly equivalent to gel detection, the use of target-specific probes in real-time PCR offers an additional level of specificity. These probes also allow the detection of single nucleotide polymorphisms (SNPs) using melt curve analysis, permitting typing of different viral species.While real-time PCR has been widely adopted for molecular infectious disease testing, real-time technology has often been compared to traditional methods such as culture, with a predictable enhancement in sensitivity (4,6,7,12). The current literature is more limited on the comparative performance of real-time PCR with established PCR formats. There is little definitive published evidence that real-time detection using fluorescent probes is more sensitive than end detection by gel, raising the question of whether real-time PCR is necessarily more sensitive than conventional PCR with gel detection. Therefore, it is essential during the validation of any real-time assay to compare its performance to something in ad...

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“…The PCR assay amplifies a 179-bp region from the HSV DNA polymerase gene (5,10,12). Samples were extracted as described above.…”

Section: Specimensmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Effect of Sequence Polymorphisms on Performance of Two Real-Time PCR Assays for Detection of Herpes Simplex Virus

Stevenson¹,

Hymas²,

Hillyard

2005

J Clin Microbiol

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“…Otro método utilizado, es el de captura híbrida, considerado como el método más confiable en la detección de 13 genotipos de alto riesgo; sin embargo, éste no permite diferenciar el tipo específico de VPH y discriminar infecciones múltiples 36,37 . La infección provocada por el VHS puede ser detectada en la actualidad mediante TAAN como la RPC-TR 38,39 . Esta técnica permite diferenciar en un solo test infecciones provocadas por VHS-1 y VHS-2, cuyos genotipos presentan una gran similitud 40 y a su vez disminuir el tiempo de detección y la cantidad de muestra utilizada considerablemente.…”

Section: Técnicas Utilizadas En La Detección De Itssunclassified

Infecciones de transmisión sexual silentes: la muestra de orina permite una adecuada detección

Retamal

Sánchez

Breb

2015

Rev. chil. infectol.

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“…Most herpesviruses have been reported to traverse the placenta (21), and intrauterine infections may cause birth defects (CMV) (19), premature delivery (VZV) (4), and fetal mortality (HSV) (21). Herpesvirus infections during pregnancy have also been associated with childhood acute lymphoblastic leukemia (EBV) (14).Common laboratory techniques in herpesvirus detection include antibody detection such as enzyme-linked immunosorbent assay (26), PCR (15,17,18,25,27), and dot blot hybridization (27). These techniques usually require separate methods for the detection of each herpesvirus and clinical information for selection of virus assay.…”

mentioning

confidence: 99%

Multiplex Detection of Human Herpesviruses from Archival Specimens by Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Sjöholm

Dillner

Carlson³

2008

J Clin Microbiol

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The human herpesviruses are involved in a variety of diseases. Large-scale evaluation of the clinical and epidemiological importance of different herpesviruses requires high-throughput methods. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a method that has a multiplex capacity enabling simultaneous detection of several viruses in a single sample. PCR-based methods for the multiplex detection of all known human herpesviruses were developed on the MALDI-TOF MS system. A variety of 882 archival samples, including bronchoalveolar lavage, conjunctival fluid, sore secretion, blister material, plasma, serum, and urine, analyzed for herpesviruses using PCR-based reference methods, were used to evaluate the MALDI-TOF MS method. The overall concordance rate between the MALDI-TOF MS method and the reference methods was 95.6% ( ‫؍‬ 0.90). In summary, the MALDI-TOF MS method is well suited for large-scale detection of all known human herpesviruses in a wide variety of archival biological specimens.The herpesviruses infecting humans include herpes simplex virus (HSV) types 1 and 2, varicella-zoster virus (VZV), Epstein-Barr virus (EBV) types A and B, cytomegalovirus (CMV), human herpesvirus 6 (HHV6) types A and B, HHV7, and HHV8 (Kaposi's sarcoma-associated herpesvirus). Herpesvirus infections in healthy subjects are often asymptomatic but can cause oral and genital lesions (HSV), chicken pox or shingles (VZV), infectious mononucleosis (EBV and CMV) (13), or roseola (HHV6) (19). In immunologically compromised hosts, herpes infections are more severe, causing pneumonia (CMV and HHV6) (1, 19), lymphoproliferative diseases (EBV) (19), Kaposi's sarcoma (HHV8) (19), or encephalitis (HSV-1) (6). Most herpesviruses have been reported to traverse the placenta (21), and intrauterine infections may cause birth defects (CMV) (19), premature delivery (VZV) (4), and fetal mortality (HSV) (21). Herpesvirus infections during pregnancy have also been associated with childhood acute lymphoblastic leukemia (EBV) (14).Common laboratory techniques in herpesvirus detection include antibody detection such as enzyme-linked immunosorbent assay (26), PCR (15,17,18,25,27), and dot blot hybridization (27). These techniques usually require separate methods for the detection of each herpesvirus and clinical information for selection of virus assay. Since most microbiological laboratories maintain large biobanks with clinical samples, large-scale evaluation of the clinical and epidemiological importance of different herpesviruses is today restricted only by cost and throughput considerations. There is therefore an increasing interest in rapid and large-scale herpesvirus detection, and methods for the multiplex detection of several herpesviruses have recently been developed using PCR and microarray techniques (8, 10, 23). Virus detection using the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system is a high-throughput technology with multiplex capacity that ha...

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Detection of Herpes simplex virus (HSV) genome using polymerase chain reaction (PCR) in clinical samples Comparison of PCR with standard laboratory methods for the detection of HSV

Cited by 53 publications

References 14 publications

Effect of Sequence Polymorphisms on Performance of Two Real-Time PCR Assays for Detection of Herpes Simplex Virus

Effect of Sequence Polymorphisms on Performance of Two Real-Time PCR Assays for Detection of Herpes Simplex Virus

Infecciones de transmisión sexual silentes: la muestra de orina permite una adecuada detección

Multiplex Detection of Human Herpesviruses from Archival Specimens by Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

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