The Identification of the Two "Missing" Romanov Children Using DNA Analysis

I first became acquainted with the Romanovs (as historical figures, not the actual Romanovs) reading in middle school about Russian History. Later, someone turned me on to Massie's Nicholas and Alexandra, which is quite a well known popular historical account of the last Czar of Russia and his family. Everyone knows the story of the end. The core of Czar's family -- the Czar Nicholas, his wife Alexandra, and his children -- had been arrested and all of them were transported to a remote location in the Urals. A complex series of events had begun involving Czarist and Revolutionary forces. At one point, it occurred to the local revolutionary officials who were responsible for the incarceration of the Czar and his family that their execution would be a good idea, to avoid their recapture by Czarists forces in the area, and to break the loyalty of Czarist supporters still resisting the Revolution. So, on July 17th, 1918, the Czar and his family, their doctor and a nurse were escorted to an empty room in the compound in which they were being held and shot. The details are rather ghastly, as summarized by an eye witness to the event.

That account does not describe a detail or two mentioned elsewhere, including the apparent fact that the Czar and his wife were shot but the daughters were killed with bayonets. I'm sure this is controversial, and the details are probably not all that important for the moment.

Anyway, after the removal of the primary iconic figures for the Czarists, the Revolution lived happily ever after.

Oh, but wait ... not so fast.

For many years, rumors persisted that one or more of the Czar's family had escaped assassination. You may remember various books, "documentaries," movies, and so on in which one or more of these individuals survives. At least one person held the claim for many years that she was the Czar's daughter, Anastasia. The most famous of these claims was made by a woman named Anna Anderson. Anderson died and was cremated in 1984, but subsequent analysis of her DNA .. on tissues stored at a hospital where she had received a medical procedure of some sort ... showed that she was not one of th Romanov girls.

But still, when the Romanov grave was eventually located and excavated, the information about that coming to light in 1991, two individuals were clearly missing. One would have been the young boy, Alexei, and the other one of the girls.

During the summer of 2007, it was reported that a second grave site had been located by an amateur archaeologist, Sergei Plotnikov. The grave included the remains of two individuals who seemed to have the necessary characteristics to have been Alexei and whichever of the girls was missing from the other grave.

Then, almost one year later, in May 2008, it was reported by Russian officials that DNA evidence linked these two individuals to the Romanov family. From a report in the New York Times:

Now an official says DNA tests have solved the mystery by identifying bone shards found in a forest as those of Aleksei and his sister Grand Duchess Maria.

...

Researchers unearthed the bone shards last summer in a forest near Yekaterinburg, where the royal family was killed, and enlisted laboratories in Russia and the United States to conduct DNA tests.

Eduard Rossel, governor of the region 900 miles east of Moscow, said Wednesday that tests done by an American laboratory had identified the shards as those of Aleksei and Maria.

"This has confirmed that indeed it is the children," he said. "We have now found the entire family."

Mr. Rossel did not specify the laboratory, but a genetic research team working at the University of Massachusetts Medical School has been involved in the process.

Now, we have a paper coming out at this very moment in PLoS ONE: Mystery Solved: The Identification of the Two Missing Romanov Children Using DNA Analysis.

Initially, it was not clear from the report what the connection might be between the reports last year and the current paper. So I contacted the authors and found out that there is a connection, though it seems a bit complicated. The present paper includes members of a large international team that has worked on the forensics in this complicated case, and this paper is the "first full detailed, forensic accounting of the the Romanov family remains." And it is quite a nice piece of work.

In this case two labs were involved, working semi-independently to replicate each other's work. Each lab successfully extracted mitochondrial DNA. Also, both labs were able to amplify low levels of nuclear DNA to get a full Y-chromosomal profile from one of the bones in the sample, confirming that individuals to be a male. The Y chromosome profile from this bone sample matched a similar profile obtained from a tooth of the Czar, confirmed by matching up with a living Romanov cousin. In addition, the labs developed a 16 marker autosomal profile for each of the individuals from both of the graves. These results clearly demonstrate that this was a family group.

Going even farther the study reports the link between the DNA sampled form the grave and DNA sampled from a shirt bloodied with the Czar's precious bodily fluids during an assassination attempt in 1891.

And so, the mystery is concluded. More or less.

Coble MD, Loreille OM, Wadhams MJ, Edson SM, Maynard K, et al. (2009). Mystery Solved: The Identification of the Two Missing Romanov Children Using DNA Analysis PLoS ONE, 4 (3)