Detection of Herpes Simplex Virus Infection in Patients With Ongoing Miscarriage Using Serological Tests and Real-Time Polymerase Chain Reaction

Background: Herpes simplex virus (HSV) is one of the most frequent viruses affecting females’ sexual and reproductive health. Objectives: The current study aimed to determine the HSV serostatus and viral shedding in patients with ongoing miscarriage. Methods: Two hundred and eight females were included in the study; IgM antibodies against HSV1/2 were detected in serum samples; the real-time polymerase chain reaction (PCR) quantification of viral DNA was performed on cervicovaginal samples. Positive females were tested for IgG anti-HSV-2. Results: The results indicated 12.5% IgM-positive and 2.9% real-time PCR positive samples. None of the patients was positive for the both analyses, simultaneously. Among IgM-positives cases, 16.6% were also IgG-positive; whilst in PCR-positives samples, 20% were also IgG-positive. The presence of viral DNA without detectable IgM or IgG antibodies could indicate a recent infection or a reactivation with low copy numbers. Conclusions: IgM alone is not a marker for viral shedding in genital tract. Molecular testing in conjunction of IgG test should be evaluated as an option to determine HSV status, and applied for research on HSV genital infections records. Keywords: Herpes, Diagnosis, Viral Sheddin

Temporal distribution and genetic variants in influenza A(H1N1)pdm09 virus circulating in Mexico, seasons 2012 and 2013

The 2012 and 2013 annual influenza epidemics in Mexico were characterized by presenting different seasonal patterns. In 2012 the A(H1N1)pdm09 virus caused a high incidence of influenza infections after a two-year period of low circulation; whereas the 2013 epidemic presented circulation of the A(H1N1)pdm09 virus throughout the year. We have characterized the molecular composition of the Hemagglutinin (HA) and Neuraminidase (NA) genes of the A(H1N1)pdm09 virus from both epidemic seasons, emphasizing the genetic characteristics of viruses isolated from Yucatan in Southern Mexico. The molecular analysis of viruses from the 2012 revealed that all viruses from Mexico were predominantly grouped in clade 7. Strikingly, the molecular characterization of viruses from 2013 revealed that viruses circulating in Yucatan were genetically different to viruses from other regions of Mexico. In fact, we identified the occurrence of two genetic variants containing relevant mutations at both the HA and NA surface antigens. There was a difference on the temporal circulation of each genetic variant, viruses containing the mutations HA-A141T / NA-N341S were detected in May, June and July; whereas viruses containing the mutations HA-S162I / NAL206S circulated in August and September. We discuss the significance of these novel genetic changes

Schematic representation of the HA molecule of the A(H1N1)pdm09 virus.

<p>HA molecules were modelled using as template the crystal structure of A 2009 H1N1 virus hemagglutinin from A/California/04/2009 (PDB 3LZG). Molecules were generated under flusurver structural model database (<a href="http://flusurver.bii.a-star.edu.sg/" target="_blank">http://flusurver.bii.a-star.edu.sg</a>). Protein sequences were blast against A/California/07/2009 to identify differences on amino acid composition. Right side HA molecule was modelled based on the strain A/Yucatan/116/2013 to indicate localization of the amino acid change S162I (red oval). The HA molecule in the middle represents the amino acidic composition of the strain A/Yucatan/81/2013 indicating the position of the mutation A141T (blue oval). Mutations at residues V234I, K283E and E499K (HA2) placed these viruses in clade 6C. Left side molecule corresponds to the HA protein sequence of the strain A/Yucatan/18/2012.</p

Amino acid changes and frequency of detection in the Neuraminidase (NA) protein of the influenza A(H1N1)pdm09 virus isolated from Yucatan, Mexico (non-Yucatan) and other regions of the world for the period 2012–2013.

<p>Percentages were calculated based on the total number of sequences analysed in this study (<i>n</i>). Empty cells indicate values = 0.</p

Temporal occurrence of genetic variants in Yucatan.

<p>Viruses from Yucatan with changes in HA and NA were distributed in time from epidemiological week 23 to 38. The bars represent the total number of influenza cases reported by the Regional Laboratory. The clear section indicates the number of confirmed cases of influenza A(H1N1)pdm09 and the grey section indicates the number of influenza A(H1N1)pdm09 viruses with genetic changes in HA and NA (HA–A141T / HA–S162I / NA–N341S / NA–L206S). The black section corresponds to the number of samples negative to influenza A(H1N1)pdm09 virus.</p

Phylogenetic tree of NA sequences of the A(H1N1)pdm09 viruses for the period 2013.

<p>Sequences from Yucatan are indicated in blue, from other regions of Mexico in red and worldwide in black. Additional amino acid changes at the node are also indicated.</p

Amino acid changes and frequency of occurrence in the hemagglutinin (HA) protein of the influenza A(H1N1)pdm09 viruses isolated in Yucatan.

<p>Frequencies of detection for each mutation are also indicated for sequences from other regions of Mexico (non-Yucatan) and worldwide for the period 2012–2013. Percentages were calculated based on the total number of sequences analysed in this study (<i>n</i>). Empty cells indicate values = 0.</p