A method for detecting rash and fever illness‐associated viruses using multiplex reverse transcription polymerase chain reaction

Abstract

In this study, a new multiplex RT-PCR method for detecting various viral genes in patients with rash and fever illnesses (RFIs) was constructed. New primer sets were designed for detection of herpes simplex viruses 1 and 2 (HSV1 and 2), and Epstein-Barr virus (EBV). The newly designed and previously reported primer sets were used to detect 13 types of RFI-associated viruses by multiplex RT-PCR assay systems. Moreover, to eliminate non-specific PCR products, a double-stranded specific DNase was used to digest double-stranded DNA derived from the templates in clinical specimens. RFI-associated viruses were detected in 77.0% of the patients (97/126 cases) by the presented method, multiple viruses being identified in 27.8% of the described cases (35/126 cases). Detected viruses and clinical diagnoses were compatible in 32.5% of the patients (41/126 cases). Sensitivity limits for these viruses were estimated to be 10 -10 copies/assay. Furthermore, non-specific PCR products were eliminated by a double-stranded specific DNase with no influence on sensitivity. These results suggest that this method can detect various RFI-associated viruses in clinical specimens with high sensitivity and specificity.

Molecular Detection of Human Herpes Viruses in Suspected Measles Serum Samples from Senegal, 2014 to 2017

“…16,17 In this report, EBV was the second most common herpes virus among the suspected cases after VZV, with 10.7% of the total number of detected viruses. This overall prevalence is consistent with reports from other studies 15,18 but lower than rates reported in previous works conducted in other geographical areas, including Mexico with 20% 19 and Germany with 16.4%. 20 Consistent with many other results, 10,16,21 the detection rates of CMV (6.6%), HHV6 (2.6%), and HSV (0.2%) were relatively low.…”

Virus detection of measles‐negative cases in Gyeonggi Province, South Korea, from 2017 to 2019

“…4,12 Furthermore, HHV7 had a 15.4% positive rate, and a similar positive rate was reported in Japan. 16 In addition, cases of HHV6 and HHV7 double infections have previously been reported as in this study. 7 The results showed that HHV6 and HHV7 increased in 2019 to 28% (28/100) and 20% (20/100), respectively.…”

“…3,4,10 Furthermore, rubella virus, PVB19, HHV6, and HHV7 have been detected in measlesnegative throat swabs and serum samples in Japan. 12,16 The results revealed that PVB19, HHV6, and HHV7 were detected in 42.2% (92/218) of measles-negative cases, except three measles-positive cases. In Japan, PVB19, the causative agent of "apple illness" with fever and rash, had a positive rate of 4.2% in measles-negative throat swabs, consistent with the results of this study.…”

“…6,11 Previous reports on PVB19 found that blood was the most commonly used test for detection, 3,10,11 although nasopharyngeal swabs have also been used. 16 HHV6 has been found in blood or bodily fluids. When the difference in positive Ct values in real-time PCR for PVB19 and HHV6 in serum and nasopharyngeal swab samples from the same patient were analyzed, it was observed that the two virus concentrations in the serum were higher than those in the nasopharyngeal swabs (data not shown).…”

“…People who had been tested for clinical signs of measles, such as fever and rash, were tested in the United States and Brazil, where measles had previously been eliminated, and, PVB19 and HHV6 were later found 3,4,10 . Furthermore, rubella virus, PVB19, HHV6, and HHV7 have been detected in measles‐negative throat swabs and serum samples in Japan 12,16 …”