Detection of hepatitis B virus DNA in liver cancer tissue by in situ polymerase chain reaction

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Detection of hepatitis E virus RNA in sera of patients with hepatitis E by polymerase chain reaction

Introduction H epatitis E, the major form of enterically transmitted non-A, non-B hepatitis, is caused by hepatitis E virus (HEV).[1] The disease is a serious public health problem in many developing countries in Asia, the Middle East, and Africa, as well as in Mexico, where sanitation conditions are suboptimal.[2] HEV is transmitted primarily by the fecal-oral route through contaminated water.[3] The overall mortality rate of hepatitis E is generally lower than 1%, but it can be as high…     

Utility of multiplex polymerase chain reaction (PCR) in diarrhea—An Indian perspective

Background

Infective diarrhea causes morbidity worldwide. Polymerase chain reaction (PCR)-based pathogen diagnostics of diarrheal stool specimens are shown to be highly sensitive and rapid as opposed to conventional diagnostics.

Methods

We analyzed the performance of FilmArray gastrointestinal (GI) panel, one such multiplex PCR test, on stool specimens in patients presenting with diarrhea to our hospital from March 2016 to September 2017 and compared the results with conventional diagnostic tests.

Results

A total of 106 patients were included. The panel detected at least one target in 54 out of 106 patients (50.9%) with results available on the same day. Multiple targets were detected in 26 out of 54 patients who tested positive (48.1%). Bacteria as an isolated etiology for diarrhea was present in 34 patients (62.9%), viruses (16.7%, nine patients), parasites (7.4%, four patients), and multiple pathogens in seven patients (12.9%). Enteroaggregative Escherichia coli (EAEC) was the commonest pathogen detected (in 23, 24% patients). Conventional diagnostic investigations, undertaken in 68/106 (64.1%) patients were positive in 12 (17.65%) as compared to 54/106 (50.9%) (p?<?0.0001). Conventional  investigations detected a pathogen not included in the study panel in 11 of 52 patients (21.1%).

Conclusion

FilmArray multiplex PCR panel detects a wide array of GI pathogens including viruses and co-infections at a shorter time with more sensitivity compared to conventional diagnostics. Henceforth, it may facilitate treatment decisions, isolation policy, and antimicrobial stewardship in patients with diarrhea requiring hospitalization.

     

Detection of influenza A virus RNA in birds by optimized Real—Time PCR system

ObjectiveTo evaluate the use of Real-Time PCR system based on specific amplification of matrix protein gene fragment for influenza A virus RNA detection in cloacal swabs from wild birds.MethodsSensitivity, specificity and reproducibility of analysis results were identified. Study of cloacal swabs from wild birds for influenza A virus presence was performed.ResultsReproducibility of low concentrations of virus detection in samples by Real-Time PCR was significantly higher than that of detection based on cytopathic effect of viruses grown on MDCK cell culture.ConclusionsReal-Time PCR system for influenza A virus RNA detection is developed and applied for virus surveillance study.     

Natural infection of Nyssomyia neivai by Leishmania (Viannia) spp. in the state of Paraná, southern Brazil, detected by multiplex polymerase chain reaction

Natural sandfly infection by Leishmania spp. in an area endemic for American cutaneous leishmaniasis was analyzed using multiplex polymerase chain reaction (PCR). The sandflies were captured using Falc?o light traps in an endemic area of the municipality of Doutor Camargo during March, April, and June 2008. In total, 1803 females were analyzed; 1755 were Nyssomyia neivai (Pinto) and 48 were Nyssomyia whitmani (Antunes and Coutinho). Multiplex PCR analyses using MP3H-MP1L and 5Llcac-3Llcac primers showed the presence of Leishmania (Viannia) spp. in 4/181 pools of sandflies, all Ny. neivai, that is, a minimal infection rate of 0.22%. This study showed, for the first time, the presence of DNA of Leishmania (Viannia) spp. in Ny. neivai. This suggests the existence of natural infection by Leishmania (Viannia) spp. in Ny. neivai in the state of Paraná. Multiplex PCR is an important tool in the detection of Leishmania infection in sandflies.     

High prevalence of Plamodium malariae infections in a Brazilian Amazon endemic area (Apiacás-Mato Grosso State) as detected by polymerase chain reaction

Plasmodium malariae is commonly confounded with Plasmodium vivax at the microscopic examination of thick blood smear. In the present study, we used a nested PCR assay to amplify a species-specific sequence of the 18S SSU rRNA gene of Plasmodium in blood samples of 497 individuals living in an endemic region of the Brazilian Amazon basin. We have found that, while the microscopic examination of thick blood smears showed a P. malariae prevalence of 1.2% (6 out of 497), the nested PCR revealed 11.9% (59 out of 497) of positive cases for this specie. These results point to the need of the development or use of a more accurate diagnosis method to distinguish between P. malariae and P. vivax, which is particularly important in view of the fact that the choice of drug for the antimalarial therapy depends on the parasite species.     

Simultaneous detection of Hb constant spring (α142, TAA>CAA, α2) and the α2 IVS-I donor site (-TGAGG) deletion by a simple polymerase chain reaction-based method in Iran

Hb Constant Spring (Hb CS, codon 142, TAA>CAA, α2) (HBA2:c.427T>C) and α2 IVS-I donor site (GAGGTGAGG>GAGG?- - - - -) (HBA2:c.95+2_95+6delTGAGG) are nondeletional α-thalassemia (α-thal) mutations found all over the world. Identification of α-thal genotypes in at-risk couples for severe anemia or in highly heterogeneous populations requires rapid, accurate and cost-effective genotyping methods. In this study, a pair of primers were used to specifically amplify an 883 bp fragment from the α2-globin gene in order to simultaneously identify these two mutations by a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method. We determined the genotypic frequencies of Hb CS and the α2 IVS-I donor site mutations after amplification and enzymatic digestion with Tru9I in 238 northern Iranian samples referred for α-thal testing. Hb CS and the α2 IVS-I donor site mutations accounted for 21 (8.8%) and 29 (12.2%) of the nondeletional cases. This genotyping assay has proven to be a rapid, reliable and useful diagnostic tool for simultaneous detection of these two anomalies for genetic counseling or further prenatal diagnosis.     

Assessing the efficacy of famotidine and rebamipide in the treatment of gastric mucosal lesions in patients receiving long-term NSAID therapy (FORCE—famotidine or rebamipide in comparison by endoscopy)

Background Nonsteroidal anti-inflammatory drugs (NSAIDs) and Helicobacter pylori infection are major causes of gastric mucosal lesions. In Japan, histamine-2 receptor antagonists are frequently prescribed, but the literature regarding their efficacy is limited. In this study, we compare the effects of famotidine and rebamipide on NSAID-associated gastric mucosal lesions using upper gastrointestinal endoscopy. Methods This study examined 112 patients taking NSAIDs for either gastric hemorrhage or erosion. Before treat-ment, the patients were assessed by endoscopy. Using blind randomization, patients were divided into two groups: group F (famotidine, 20 mg/day) and group R (rebamipide, 300 mg/day). Efficacy was examined 4 weeks later using endoscopy. Results After treatment, the Lanza score decreased significantly in group F (P < 0.001) but not in group R (P = 0.478). The change in the Lanza score in group F was significantly greater (P = 0.002) than that in group R. Conclusions Famotidine was superior to rebamipide in treating NSAID-associated mucosal lesions.     

The first determination of Eustrongylides excisus Jägerskiöld, 1909 — larvae (Nematoda: Dioctophymatidae) in the pike-perch Sander lucioperca in Vojvodina (Serbia)

Twenty-one specimens of pike-perch (Sander lucioperca) were caught in the Danube-Tisa-Danube Canal in the city area of Novi Sad for parasitological examination. The presence of nematodes in the muscles was revealed in three fish. The parasites were identified to belong to the species Eustrongylides excisus, for which the pike-perch is a paratenic host. This finding represents the first determination of the larvae in the pike-perch in Serbia. The pike-perch is infected by ingestion of benthos- or plankton-eating fishes, the second intermediate hosts harbouring the fourth-stage nematode larvae. E. excisus is pathogenic to humans, who may be infected by consuming raw or undercooked fish.     

Appropriateness in prescribing PPIs: A position paper of the Italian Society of Gastroenterology (SIGE) — Study section “Digestive Diseases in Primary Care”

The introduction of proton pump inhibitors (PPIs) into clinical practice about thirty years ago has greatly improved our therapeutic approach to acid-related diseases for their well-recognized efficacy and safety.Despite the well-defined indications, however, the use of PPIs continues to grow every year in both western and eastern countries and this phenomenon poses serious queries that include the onset of potential adverse effects and the increase in health care costs. The major reason explaining this worrying market expansion is the inappropriate use of PPIs. In order to re-establish a correct use of these effective drugs in daily clinical practice, the Italian Society of Gastroenterology (SIGE), nominated a panel of experts who reviewed the available clinical literature and produced a series of updated position statements on the use of PPIs in clinical practice.