Co-occurrence of Herpes Simplex Virus 1 and 2 in Patients Suspected with Different Neurological Ailments in Karachi

Consensus PCR assays that can be used to sensitively detect several herpesvirus (HV) species across the different subfamilies were developed in this study. Primers containing degenerate bases were designed to amplify regions of the DNA polymerase (DPOL) gene of alpha- and gamma-HVs, and the glycoprotein B (gB) gene of beta-HVs in a singleplex, non-nested touchdown PCR format. The singleplex touchdown consensus PCR (STC-PCR) was used to amplify the DNA of eight human and 24 animal HVs. The assay was able to detect the lowest DNA dilution of 10−5 for alpha-HVs and 10−3 for beta- and gamma-HVs. In comparison, lowest detection limits of 10−5, 10−3, and 10−2 were obtained for alpha-, beta-, and gamma-HVs respectively when a nested PCR was used. The findings in this study suggest that the STC-PCR assays can be employed for the molecular surveys and clinical detection of novel and known HVs.

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Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses

Okoh

Lockhart

Grimsey

et al. 2023

Eur J Clin Microbiol Infect Dis

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Metagenomic search of viral coinfections in herpes simplex encephalitis patients

Perlejewski,

Radkowski,

Rydzanicz

et al. 2023

J. Neurovirol.

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Little is known about concomitant central nervous system (CNS) infections by more than one virus. Current diagnostics are based on molecular tests for particular pathogens making it difficult to identify multi-viral infections. In the present study, we applied DNA- and RNA-based next-generation sequencing metagenomics (mNGS) to detect viruses in cerebrospinal fluids from 20 patients with herpes simplex encephalitis. Coinfection was detected in one patient: sequences in cerebrospinal fluids matched enterovirus A (2.660 reads; 4% of recovered genome) and enterovirus B (1.571 reads; 13% of recovered genome). Subsequent PCR combined with serotyping allowed to identify human echovirus 6, a representative of enterovirus B. Several other mNGS hits (human pegivirus, Merkel cell polyomavirus, human papillomavirus type 5) were not considered to represent a genuine signal as they could not be confirmed by specific RT-PCR/PCR. HSV DNA, while being detectable by PCR in every patient, was detected by mNGS in only one. In conclusion, contaminations and false signals may complicate mNGS interpretation; however, the method can be useful in diagnostics of viral coinfections in CNS, particularly in the case of rare pathogens.

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Co-occurrence of Herpes Simplex Virus 1 and 2 in Patients Suspected with Different Neurological Ailments in Karachi

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Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses

Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses

Metagenomic search of viral coinfections in herpes simplex encephalitis patients

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