DNA amplification by polymerase chain reaction (PCR) should be inhibited if the target for amplification region in the template DNA is nicked or cut. Based on this premise, we established a sensitive and differential assay using PCR to detect antibiotics that act on DNA. After template lambda DNA (10 pg) was incubated with antibiotics (10 approximately 20 ng) at 37 degrees C for 30 minutes in a...

We have developed a web-enabled system called MuPlex that aids researchers in the design of multiplex PCR assays. Multiplex PCR is a key technology for an endless list of applications, including detecting infectious microorganisms, whole-genome sequencing and closure, forensic analysis and for enabling flexible yet low-cost genotyping. However, the design of a multiplex PCR assays is computatio...

Detection of Coxiella burnetii, the etiologic agent of Q fever, is important for diagnosis of Q fever. PCR-based methods have been widely used for the detection mostly because isolation of C. bumetii is time-consuming. Recent reports showed that PCR-positive rates of Q fever infection widely differed. We have evaluated the PCR and nested PCR assays currently used in Japan. The nested PCR assay ...

An enrichment broth culture-duplex PCR combination assay was devised to identify Clostridium perfringens directly from fecal samples. The method consists of a combination of short enrichment of samples in selective media, DNA isolation, and performing duplex PCR using two pairs of primers which identify C. perfringens strains that harbor the virulence enterotoxin gene. Comparison of two selecti...

We have read with interest the recent article by Morata et al. reporting their study of two of the possible factors which can interfere with specific DNA amplification in a peripheralblood PCR assay used for the diagnosis of human brucellosis (2). The study deals with the optimization of a method described in a previous work by these authors (4), in which they report a sensitivity of 100% and a...

The presence of PCR inhibitors in water samples is well known and contributes to the fact that a practical PCR assay has not been developed for legionella surveillance. In this study, we devised a new seminested PCR assay for detection of Legionella spp. in water samples as a means of overriding the PCR inhibitors without loss of sensitivity. The seminested PCR assay utilized primers to amplify...

This is the first published report of a PCR assay for detecting porcine cytomegalovirus (PCMV), the causative agent of inclusion body rhinitis in pigs. The DNA to be tested was extracted directly from lungs and nasal scrapings of pigs with various clinical syndromes. Fifty-nine percent (74 of 126) of tested pigs with various clinical syndromes were found to be PCR positive for PCMV. It is hoped...

background: limiting dilution assay is considered as the gold standard method for quantifying the number of parasites in the animal model of leishmania infection. nowadays, real-time pcr is being increasingly applied to quantify infectious agents. in the present study , a real-time pcr assay was developed to estimate para­site burdens in lymph nodes of leishmania major infected balb/c mice. enu...

background: acinetobacter baumannii is an important opportunistic pathogen which causes complications in hospitalized patients, especially those in icu. the aim of this study was to determine the frequency of class 1 and 2 integrons in multi-drug resistance a. baumannii and to investigate the association between the presence of integrons and antibiotic resistance patterns. methods: a total of 4...