A year or so ago, Nature Communications published an article that highlighted how researchers in the Department of Genetics at the University of Leicester successfully used DNA from saliva to help identify the remains of King Richard III and solved a 500 year old mystery^[1]. The story behind the recovery of the remains and their subsequent identification as those of the missing king is an interesting one that combines ancient DNA analysis with modern DNA from saliva.

First, let’s look at the background of King Richard. Richard III was the last English king to be killed in battle and his precise resting place was not known. According to the article in Nature Communications^[1]:

“Historical records report that after Richard III was killed on the battlefield, age 32, his remains were brought back to Leicester and buried in the medieval church of the Grey Friars. The friary was dissolved in 1538 under the orders of King Henry VIII..... the exact site of Richard III’s grave had been lost in the 527 years since his death.”

However, in September of 2012, a team of investigators searched the area thought to be the site of the friary and excavated a skeleton. The skeletal evidence was consistent with the remains being those of Richard III; (male aged 30-34 years, severe scoliosis, and battle injuries matching those King Richard was expected to have). What was missing was genetic evidence to support the non-genetic data to confirm identification.

Researchers at the University of Leicester took on the task to confirm the identity using DNA. They knew that DNA identification would rely on a comparison of the DNA from the skeleton compared to DNA from known living relatives. The challenge was to find potential relatives to take part in the study. The researchers were able to locate and contact 5 male-line relatives and two female-line relatives who agreed to participate in the study.

DNA was extracted from teeth and the femur for the skeleton with duplicate analysis of the DNA being conducted in separate ancient DNA laboratories to replicate results. DNA was extracted from saliva samples of the living relatives. DNA from these samples was sequenced twice in both the forward and reverse direction with no differences found between replicates or between samples.

After reviewing all the available evidence from the DNA analysis of the ancient skeleton and identified modern relatives using highly conservative measures, the researchers concluded that:

“…the evidence is overwhelming that Skeleton 1 from the Grey Friars site in Leicester is that of Richard III, thereby closing a 500-year old plus missing person case”.

The story does not end there. In March of 2015, King Richard III was reinterred in Leicester with all the pomp and grandeur of a state funeral. The event was attended by modern day royalty, military and the academic elite of Britain. The resting place of this King is no longer a mystery, thanks to ancient and modern DNA analysis (using Oragene/saliva collection). 

References:

[1] King, T. E. et al. Identification of the remains of King Richard III. Nat. Commun. 5:5631 doi: 10.1038/ncomms6631 (2014).