Restriction site insertion-PCR (RSI-PCR) for rapid discrimination and typing of closely related microbial strains

Restriction site-free insertion of PCR products directionally into vectors.

A restriction site-free cloning method has been developed for inserting a PCR product into a vector flexibly and precisely at any desired location with high efficiency. The method uses a pair of DNA integration primers with two portions. The 3' portion isolates the inserts by PCR, and the 5' portion integrates the PCR products into the homologous region of the vector. For mutagenesis, a third p...

Discrimination of SHV beta-lactamase genes by restriction site insertion-PCR.

Restriction site insertion-PCR (RSI-PCR) is a simple, rapid technique for detection of point mutations. This technique exploits primers with one to three base mismatches near the 3' end to modulate a restriction site. We have developed this technique to identify described mutations of the bla(SHV) genes for differentiation of SHV variants that cannot be distinguished easily by other techniques....

PCR and RFLP Analysis for Identification and Typing of Helicobacter pylori Strains from Gastric Biopsy Specimens

SmaI restriction site-based multiplex PCR for typing of hospital- and community-acquired Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen, and morbidity and mortality rates associated with this pathogen have increased markedly in recent years. MRSA strains are generally resistant to several classes of antibiotics and are therefore difficult and costly to treat. A major issue is to identify the sources of MRSA infections and to monitor their epi...

Discrimination of Mycobacterium tuberculosis strains by PCR.