In 1998, when the PCR technique was already popular, a Japanese company called Eiken Chemical Co., Ltd. designed a method known as the loop-mediated isothermal amplification of DNA (LAMP). The method can produce up to 109 copies of the amplified DNA within less than an hour. It is also highly specific due to the use of two to three pairs of primers (internal, external, and loop), which recognise up to eight specific locations on the DNA or RNA targets. Furthermore, the Bst DNA polymerase most used in LAMP shows a high strand displacement activity, which eliminates the DNA denaturation stage. One of the most significant advantages of LAMP is that it can be conducted at a stable temperature, for instance, in a dry block heater or an incubator. The products of LAMP can be detected much faster than in standard techniques, sometimes only requiring analysis with the naked eye. The following overview highlights the usefulness of LAMP and its effectiveness in various fields; it also considers the superiority of LAMP over PCR and presents RT-LAMP as a rapid diagnostic tool for SARS-CoV-2.
Recent investigations of East Asian freshwater mussel genera Nodularia, Middendorffinaia and Inversiunio confirmed validity of the taxa, while their composition and taxonomic uncertainty concerning classification of these genera within a specific subfamily are still in question. The aim of the present study is to clarify the taxonomy and systematics of Nodularia and Middendorffinaia genera from the Russian Far East and Inversiunio from Japan, using a multiple dataset approach that combines morphology of glochidia, detailed conchological analyses with morphometry of adult and glochidial shells, and a molecular technique involving COI barcoding marker. As a result, three genetically distinct and highly divergent lineages were recognized confirming the existence of three distinct genera. The existence of five Nodularia species in Southeast Asia is shown. One of the sequences is shown to be incorrectly assigned to N. jourdyi, while in fact it also represents N. douglasiae, according to the BIN system. Existence of a single Nodularia species, namely N. douglasiae, and a single Middendorffinaia species, namely M. mongolica, in the Russian Far East is supported. We conclude that so-called comparatory species Nodularia amurensis, N. schrencki, and N. vladivostokensis are junior synonyms of the species Nodularia douglasiae; the comparatory species Middendorffinaia ussuriensis and M. dulkeitiana are junior synonyms of the species Middendorffinaia mongolica. Within Inversiunio specimens from Japan four species were recognized based on types of glochidial microsculpture of shells and genetic data. Gene sequences of Inversiunio verrucosus, endemic to the Seomjingang River basin in South Korea, and I. yanagawensis from Kyushu Island in Japan are very similar and they can be regarded as the same species, with Inversiunio verrucosus as a junior synonym of Inversiunio yanagawensis. Mussels from Honshu Island first referred to I. yanagawensis are shown to be a separate species.
The alien Chinese mussel Anodonta woodiana was first reported in Poland in the system of heated lakes near Konin in 1993. Genetic studies with use of three molecular techniques (isoenzyme electrophoresis, PCR-RFLP and sequence analysis of a COI gene fragment) were carried out on the Polish first populations of A. woodiana. The studies have revealed low genetic variation between the populations (Nei's genetic distance for 12 loci ranged 0.000 to 0.007) as well as their considerable polymorphism. Each population averaged 2.28 alleles per locus, 2.72 alleles per polymorphic locus, and 75% polymorphic loci. Restriction analysis of the COI gene fragment have not revealed variability between the analysed specimens, including males and females. Restriction enzymes, ScrFI, Csp6I, and EcoRI used in the COI gene fragment PCR-RFLP generate distinct restriction patterns, which can be molecular markers for A. woodiana. The sequence obtained for COI fragment was the same in the examined female and male specimens and represents F mitotype (DNA was isolated from somatic tissues). The divergence between A. woodiana F and M mitotypes is high (34%), however it remains within the range of the general character of the DUI (doubly uniparental inheritance) phenomenon in freshwater bivalves (Unionidae).