[Establishment of loop-mediated isothermal amplification method for detection of Legionella pneumophila].

Objective: To establish a simple and rapid molecular detection for Legionella pneumophila.

Methods: The loop-mediated isothermal amplification (LAMP) was applied for detection of Legionella pneumophila. A set of primers were designed to identify six special areas in mip gene of Legionella pneumophila.

[Analysis of natural killer cell immunoglobulin-like receptor gene family in HLA-identical sibling].

The aim of study was to analyze natural killer immunoglobulin (Ig)-like receptor (KIR) gene content in HLA-identical sibling and to investigate the possibility of their KIR match. Samples were genotyped for HLA by Luminex method and polymerase chain reaction sequence based typing, the KIR gene was detected by polymerase chain reaction sequence-specific primers. The results showed that 17 KIR genes could be observed in the 27 pairs HLA-A, -B, -Cw and -DRB1 locus identical sibling samples.

[Sequence analysis of a novel HLA-A * 2459 allele].

This study was aimed to investigate the molecular genetics basis of a novel allele HLA-A * 2459 in Chinese population. DNA was extracted from whole blood by PEL-FREEZ DNA extraction kit. The amplification of HLA-A exons 2 - 4 of the proband was preformed by allele specific primer PCR and the amplified product was sequenced bidirectionally with primers.

[Identification and sequence analysis of a null HLA-B allele HLA-B*5408N newly detected].

The study was purposed to investigate the molecular genetic basis for HLA novel allele HLA-B*5408N in Chinese population. DNA was extracted from whole blood by commercial DNA extraction kit, the HLA-B exons 2 - 4 of the proband was amplified by allele specific primers PCR and the amplified product was sequenced for exons 2, 3 and 4 bidirectionally. The sequencing results showed HLA-B alleles of the proband as B*1527 and the novel allele.

[Sequence analysis of HLA-B*4061 allele newly found].

The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B*4061, in Chinese population. DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart.

[Allelic polymorphism analysis of killer cell immunoglobulin-like receptor gene 3DL2 in Zhejiang Han population].

Objective: To analyze the allelic polymorphism of killer cell immunoglobulin (Ig)-like receptor (KIR) gene 3DL2 in Zhejiang Han population.

Methods: Allelic definition within KIR3DL2 was achieved through using the PCR-sequence specific oligonucleotide probes (PCR-SSOP) method and the specific amplification of exon 3-exon 4 and exon 8-exon 9. Twenty-one digoxigenin-labelled probes were used to the SSOP analysis.

[Establishment of delta block matching technique].

To establish delta block HLA-matching technique, DNA was extracted from whole blood by salting-out method, delta block was amplified by polymerase chain reaction (PCR), and PCR product was detected by GeneScan. The results showed that delta block had polymorphism in 104 samples without sibship of the Han people from Zhejiang province. The range of DNA fragment length was 81-393 bp and could be divided into 4 groups: 81-118 bp, 140-175 bp, 217-301 bp, 340-393 bp.

[Sequence analysis of a novel HLA-DRB1 allele, DRB1 * 1212].

Objective: To investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.

Methods: Genomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA.

[Polymorphism of killer cell immunoglobulin-like receptors gene family in Zhejiang Han population].

To analyze killer immunoglobulin (Ig)-like receptor (KIR) gene content and allelic polymorphism in Zhejiang Han population, samples were genotyped by polymerase chain reaction sequence-specific primers (PCR-SSP). The results demonstrated that all 17 KIR genes could be observed in the population. All individuals contained 2DL4, 3DL2 and 3DL3 genes.

[Establishment of beta block matching technique].

The purpose of this study was to establish beta block matching technique. DNA was extracted from whole blood by salting-out method, beta block matching was performed by PCR and GeneScan technique. The results showed that the length of fragments amplificated in 100 samples was different and the range of them was 91-197 bp.

[Identification and sequence analysis of a novel HLA-B*5614 allele].

Objective: To investigate the molecular genetics basis for a novel HLA allele, HLA-B*5614, in Chinese population.

Methods: DNA was extracted from whole blood by salting-out method. The HLA-B exons 2-4 of the proband was amplified and the amplified product was cloned using TOPO cloning sequencing kit to split the two alleles apart.